Cloning and Genetic Modification (GM/GMO's)

ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) I. SUB-EXHIBIT INFORMATION ABOUT CELLS AND GENETICS At a microscopic level, we are all composed of cells. Look at yourself in a mirror -- what you see is about 10 trillion cells divided into about 200 different types. Our muscles are made of muscle cells, our livers of liver cells, and there are even very specialized types of cells that make the enamel for our teeth or the clear lenses in our eyes! If you want to understand how your body works, you need to understand cells. Everything from reproduction to infections to repairing a broken bone happens down at the cellular level. If you want to understand new frontiers like biotechnology and genetic engineering, you need to understand cells as well. A. CELLS The cell is the basic structural, functional and biological unit of all known living organism. Cells are the smallest unit of life that is classified as a living thing, and are often called the "building blocks of life". The word cell comes from the Latin cella, meaning "small room". It was coined by Robert Hooke in his book Micrographia (1665), in which he compared the cork cells he saw through his microscope to the small rooms monks lived in. The cell was discovered by Robert Hooke in 1665. The cell theory, first developed in 1839 by Matthias Jakob Schleiden and Theodor Schwann, states that all organisms are composed of one or more cells, that all cells come from preexisting cells, that vital functions of an organism occur within cells, and that all cells contain the hereditary information necessary for regulating cell functions and for transmitting information to the next generation of cells. Cells emerged on Earth at least 3.5 billion years ago. Cells consist of protoplasm enclosed within a membrane, which contains many biomolecules such as proteins and nucleic acids. Organisms can be classified as unicellular (consisting of a single cell; including most bacteria) or multicellular (including plants and animals). While the number of cells in plants and animals varies from species to species, humans contain about 100 trillion (1014) cells. Most plant and animal cells are between 1 and 100 micrometres and therefore are visible only under the microscope. 1 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) There are two types of cells, eukaryotes, which contain a nucleus, and prokaryotes, which do not. Prokaryotic cells are usually single-celled organisms, while eukaryotic cells can be either single-celled or part of multicellular organisms. Prokaryotic cells were the first form of life on Earth. They are simpler and smaller than eukaryotic cells, and lack membrane-bound organelles such as the nucleus. Prokaryotes include two of the domains of life, bacteria and archaea. The DNA of a prokaryotic cell consists of a single chromosome that is in direct contact with the cytoplasm. The nuclear region in the cytoplasm is called the nucleoid. Plants, animals, fungi, slime moulds, protozoa, and algae are all eukaryotic. These cells are about fifteen times wider than a typical prokaryote and can be as much as a thousand times greater in volume. The main distinguishing feature of eukaryotes as compared to prokaryotes is compartmentalization: the presence of membrane-bound compartments in which specific metabolic activities take place. Most important among these is a cell nucleus, a membrane-delineated compartment that houses the eukaryotic cell's DNA. This nucleus gives the eukaryote its name, which means "true nucleus." Figure 1.0 Structure of a plant cell. Figure 1.1 Structure of an animal cell. 2 Coming Soon ONE ACTION. ONE MILLION CONSEQUENCES. (Cloning and GMOs) For a layman, the primary difference between plants and animals is that the former remains fixed, while the latter has the ability to move themselves from one place to another. But, there is more to this that differentiates a plant from an animal, in terms of their cell anatomical structure and parts. Listed below are some of the distinguishing features between a plant cell and an animal cell. Animal Cell Plant Cell Cell wall: Absent Present (formed cellulose) Shape: Round (irregular shape) Rectangular shape) Vacuole: One or more small vacuoles (much smaller than plant cells). One, large central vacuole taking up 90% of cell volume. Centrioles: Present in all animal cells Only present in lower plant forms. Chloroplast: Animal cells don't have chloroplasts Plant cells have chloroplasts because they make their own food Cytoplasm: Present Present Endoplasmic Reticulum (Smooth and Rough): Present Present Ribosomes: Present Present of (fixed 3 Coming Soon ONE ACTION. ONE MILLION CONSEQUENCES. (Cloning and GMOs) B. Animal Cell Plant Cell Mitochondria: Present Present Plastids: Absent Present Golgi Apparatus: Present Present Plasma Membrane: only cell membrane cell wall and membrane Microtubules/Microfilaments: Present Present Flagella: May be found in some cells May be found in some cells Lysosomes: Lysosomes cytoplasm. Lysosomes usually not evident. Nucleus: Present Present Cilia: Present It is very rare occur in a cell INSIDE THE NUCLEUS The nucleus is a highly specialized organelle that serves as the information processing and administrative center of the cell. This organelle has two major functions: it stores the cell's hereditary material, or DNA, and it coordinates the cell's activities, which include growth, intermediary metabolism, protein synthesis, and reproduction (cell division). 4 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) Only the cells of advanced organisms, known as eukaryotes, have a nucleus. Generally there is only one nucleus per cell, but there are exceptions, such as the cells of slime molds and the Siphonales group of algae. Simpler one-celled organisms (prokaryotes), like the bacteria and cyanobacteria, don't have a nucleus. In these organisms, all of the cell's information and administrative functions are dispersed throughout the cytoplasm. The spherical nucleus typically occupies about 10 percent of a eukaryotic cell's volume, making it one of the cell's most prominent features. A double-layered membrane, the nuclear envelope, separates the contents of the nucleus from the cellular cytoplasm. The envelope is riddled with holes called nuclear pores that allow specific types and sizes of molecules to pass back and forth between the nucleus and the cytoplasm. It is also attached to a network of tubules and sacs, called the endoplasmic reticulum, where protein synthesis occurs, and is usually studded with ribosomes. The semifluid matrix found inside the nucleus is called nucleoplasm. Within the nucleoplasm, most of the nuclear material consists of chromatin, the less condensed form of the cell's DNA that organizes to form chromosomes during mitosis or cell division. The nucleus also contains one or more nucleoli, organelles that synthesize protein-producing macromolecular assemblies called ribosomes, and a variety of other smaller components, such as Cajal bodies, GEMS (Gemini of coiled bodies), and interchromatin granule clusters. Chromatin and Chromosomes - Packed inside the nucleus of every human cell is nearly 6 feet of DNA, which is divided into 46 individual molecules, one for each chromosome and each about 1.5 inches long. Packing all this material into a microscopic cell nucleus is an extraordinary feat of packaging. For DNA to function, it can't be crammed into the nucleus like a ball of string. Instead, it is combined with proteins and organized into a precise, compact structure, a dense string-like fiber called chromatin. The Nucleolus - The nucleolus is a membrane-less organelle within the nucleus that manufactures ribosomes, the cell's protein-producing structures. Through the microscope, the nucleolus looks like a large dark spot within the nucleus. A nucleus may contain up to four nucleoli, but within each species the number of nucleoli is fixed. After a cell divides, a nucleolus is formed when chromosomes are brought together into nucleolar organizing 5 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) regions. During cell division, the nucleolus disappears. Some studies suggest that the nucleolus may be involved with cellular aging and, therefore, may affect the senescence of an organism. The Nuclear Envelope - The nuclear envelope is a double-layered membrane that encloses the contents of the nucleus during most of the cell's lifecycle. The space between the layers is called the perinuclear space and appears to connect with the rough endoplasmic reticulum. The envelope is perforated with tiny holes called nuclear pores. These pores regulate the passage of molecules between the nucleus and cytoplasm, permitting some to pass through the membrane, but not others. The inner surface has a protein lining called the nuclear lamina, which binds to chromatin and other nuclear components. During mitosis, or cell division, the nuclear envelope disintegrates, but reforms as the two cells complete their formation and the chromatin begins to unravel and disperse. Nuclear Pores - The nuclear envelope is perforated with holes called nuclear pores. These pores regulate the passage of molecules between the nucleus and cytoplasm, permitting some to pass through the membrane, but not others. Building blocks for building DNA and RNA are allowed into the nucleus as well as molecules that provide the energy for constructing genetic material. C. DNA DNA, short for deoxyribonucleic acid, is the molecule that contains the genetic code of organisms. This includes animals, plants, protists, archaea and bacteria. DNA is in each cell in the organism and tells cells what proteins to make. A cell's proteins determine its function. DNA is inherited by children from their parents. This is why children share traits with their parents, such as skin, hair and eye color. The DNA in a person is a combination of the DNA from each of their parents. DNA was first isolated (extracted from cells) by Swiss physician Friedrich Miescher in 1869, when he was working on bacteria from the pus in surgical bandages. The molecule was found in the nucleus of the cells and so he called it nuclein. DNA's role in heredity was confirmed in 1952, when Alfred Hershey and Martha Chase in the Hershey–Chase experiment showed that DNA is the genetic material of the T2 bacteriophage. 6 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) In 1953, James D. Watson and Francis Crick suggested what is now accepted as the first correct double-helix model of DNA structure in the journal Nature. Their double-helix, molecular model of DNA was then based on a single X-ray diffraction image "Photo 51", taken by Rosalind Franklin and Raymond Gosling in May 1952. How Watson and Crick got Franklin's results has been much debated. Crick, Watson and Maurice Wilkins were awarded the Nobel Prize in 1962 for their work on DNA. DNA is found inside a special area of the cell called the nucleus. Because the cell is very small, and because organisms have many DNA molecules per cell, each DNA molecule must be tightly packaged. This packaged form of the DNA is called a chromosome. During DNA replication, DNA unwinds so it can be copied. At other times in the cell cycle, DNA also unwinds so that its instructions can be used to make proteins and for other biological processes. But during cell division, DNA is in its compact chromosome form to enable transfer to new cells. DNA is made of chemical building blocks called nucleotides. These building blocks are made of three parts: a phosphate group, a sugar group and one of four types of nitrogen bases. To form a strand of DNA, nucleotides are linked into chains, with the phosphate and sugar groups alternating. The four types of nitrogen bases found in nucleotides are: adenine (A), , thymine (T), guanine (G) and cytosine (C). The order, or sequence, of these bases determines what biological instructions are contained in a strand of DNA. For example, the sequence ATCGTT might instruct for blue eyes, while ATCGCT might instruct for brown. Each DNA sequence that contains instructions to make a protein is known as a gene. The size of a gene may vary greatly, ranging from about 1,000 bases to 1 million bases in humans. The complete DNA instruction book, or genome, for a human contains about 3 billion bases and about 20,000 genes on 23 pairs of chromosomes. 7 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) D. HEREDITY Heredity is the transmission of genetic characteristics from ancestor to descendant through the genes. As a subject, it is tied closely to genetics, the area of biological study concerned with hereditary traits. The study of heritable traits helps scientists discern which are dominant and therefore are likely to be passed on from one parent to the next generation. On the other hand, a recessive trait will be passed on only if both parents possess it. Among the possible heritable traits are genetic disorders, but study in this area is ongoing, and may yield many surprises. The idea of particulate inheritance of genes can be attributed to the Moravian monk Gregor Mendel who published his work on pea plants in 1865. However, his work was not widely known and was rediscovered in 1901. It was initially assumed the Mendelian inheritance only accounted for large (qualitative) differences, such as those seen by Mendel in his pea plants—and the idea of additive effect of (quantitative) genes was not realised until R. A. Fisher's (1918) paper, "The Correlation Between Relatives on the Supposition of Mendelian Inheritance" Mendel's overall contribution gave scientists a useful overview that traits were inheritable. As of today, his pea plant demonstration became the foundation of the study of Mendelian Traits. These traits can be traced on a single locus. In humans, eye color is an example of an inherited characteristic: an individual might inherit the "brown-eye trait" from one of the parents. Inherited traits are controlled by genes and the complete set of genes within an organism's genome is called its genotype. The complete set of observable traits of the structure and behavior of an organism is called its phenotype. These traits arise from the interaction of its genotype with the environment. As a result, many aspects of an organism's phenotype are not inherited. For example, suntanned skin comes from the interaction between a person's phenotype and 8 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) sunlight; thus, suntans are not passed on to people's children. However, some people tan more easily than others, due to differences in their genotype: a striking example is people with the inherited trait of albinism, who do not tan at all and are very sensitive to sunburn. Heritable traits are known to be passed from one generation to the next via DNA, a molecule that encodes genetic information. DNA is a long polymer that incorporates four types of bases, which are interchangeable. The sequence of bases along a particular DNA molecule specifies the genetic information: this is comparable to a sequence of letters spelling out a passage of text. Before a cell divides through Mitosis, the DNA is copied, so that each of the resulting two cells will inherit the DNA sequence. A portion of a DNA molecule that specifies a single functional unit is called a gene; different genes have different sequences of bases. Within cells, the long strands of DNA form condensed structures called chromosomes. The specific location of a DNA sequence within a chromosome is known as a locus. If the DNA sequence at a particular locus varies between individuals, the different forms of this sequence are called alleles. DNA sequences can change through mutations, producing new alleles. If a mutation occurs within a gene, the new allele may affect the trait that the gene controls, altering the phenotype of the organism. However, while this simple correspondence between an allele and a trait works in some cases, most traits are more complex and are controlled by multiple interacting genes within and among organisms. Developmental biologists suggest that complex interactions in genetic networks and communication among cells can lead to heritable variations that may underlay some of the mechanics in developmental plasticity and canalization. Recent findings have confirmed important examples of heritable changes that cannot be explained by direct agency of the DNA molecule. These phenomena are classed as epigenetic inheritance systems that are causally or independently evolving over genes. Research into modes and mechanisms of epigenetic inheritance is still in its scientific infancy, however, this area of research has attracted much recent activity as it broadens the scope of heritability and evolutionary biology in general. DNA methylation marking chromatin, self-sustaining metabolic loops, gene silencing by RNA interference, and the three dimensional conformation of proteins (such as prions) are areas where epigenetic inheritance systems have been discovered at the organismic level. Heritability may also occur at even larger scales. For example, ecological inheritance through the process of niche construction is defined by the regular and repeated activities of organisms in their environment. This generates a 9 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) legacy of effect that modifies and feeds back into the selection regime of subsequent generations. Descendants inherit genes plus environmental characteristics generated by the ecological actions of ancestors. Other examples of heritability in evolution that are not under the direct control of genes include the inheritance of cultural traits, group heritability, and symbiogenesis. These examples of heritability that operate above the gene are covered broadly under the title of multilevel or hierarchical selection, which has been a subject of intense debate in the history of evolutionary science. II. Main Exhibit A. Cloning and GMOs Seen in the Media For Cloning: Popular sci-fi movies that feature cloning include Jurassic Park, The Island and the like. Jurassic Park Cloning can be done using a single drop of blood which contains billions strands of DNA (the building blocks of life). DNA strand is a blue print of building a living thing. A full DNA strand contains 3 billion genetic codes. Cloning was accomplished by extracting the DNA of dinosaurs from mosquitoes that had been preserved inside fossilized amber. Amber is fossilized tree resin. However, the strands of DNA were incomplete, so DNA from frogs was used fill in the gaps to produce dinosaur eggs. The dinosaurs all were cloned genetically as females in order to prevent breeding. But because they have the genetic coding of frog DNA - West African bullfrogs which can change their gender in a single-sex environment, in which the cloned dinosaurs were able to do as well. These huge advancements in scientific technology have enabled a mogul to create an island full of living dinosaurs for a park that was built with genetically engineered dinosaurs. The Island Dr. Merrick runs the Merrick Institute, a bio-engineering facility where the Agnates are grown. An Agnate is a clone of a regular person; grown directly into adulthood, matching the biological age of the client; its DNA completely identical to the client's. Dr. Merrick falsely claims that the Agnates are kept in a vegetative state, never achieving consciousness, and never thinking or feeling, in full compliance with eugenics laws of 2015. An Agnate is meant as a source of replacement body parts for an ailing client; each Agnate being a perfect DNA match for its client, there is never worry about rejection of body parts, 10 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) nor a need to wait for available organs, during which time the client could die. Female Agnates can also serve as surrogate wombs for a female client who cannot carry a baby to term, herself (Lima One Alpha). He stresses his false claim that Agnates do not achieve sentience, and are products; not human, as humans think of themselves as human. Matanglawin Matanglawin featured cloning. Cloning was explained as a way of science where the act of copying an organism with the exact traits, appearance and behavior using genetics. Cloning can be done using somatic cell nuclear transfer. Each organism consists of cells and in each cell contains the nucleus which has the genes of any species. It is like an identification card that holds information like the color of eyes, hair, height and any other personal qualities. The nucleus can be acquire and transferred to an egg cell. It is possible to produce an offspring which have the exact quality of the nucleus used. Cloning can be done to animal whose have great features like cows. Matanglawin also explained the movie Jurassic Park. The show also exampled a cloned cat called CC and the possible cloning of a best preserved mammoth named Yuka . They also exampled cloning projects in the Philippines like ripening of mangoes and papaya by genetic engineering and increasing population of endangered species of trees. Matanglawin discussed brief informations about human cloning. For GMOs: Experts in GMOs discussed videos in Youtube. That says 90% crops and many other products that came from US are genetically modified foods. These include fast food chains like Mcdo, Pizza Hut and the like. For Cloning and GMOs: Bato Balani Bato Balani is a science magazine that has been helping the Filipino youth for over 25 years by sharing relevant and significant information in the field of science and technology. They have made articles about cloning and GMOs. 11 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) B. Brief History 1973 in Honolulu, Hawaii, Herbert Boyer (left) and Stanley Cohen (right) combined their efforts in biotechnology to invent a method of cloning genetically engineered molecules in foreign cells. It is a technique of DNA cloning, which allowed genes to be transplanted between different biological species. (Using Boyer's methodology, they were able to successfully introduce foreign DNA into bacterial plasma, and using Cohen's methodology, they were able to subsequently insert this modified plasmid into bacteria. Because bacteria divide very rapidly, their work allowed the genetic "manufacturing" of engineered drugs and hormones, leading to the multi-billion dollar biotechnology industry.) Identification of the Ti plasmid in a bacteria (Agrobacterium tumefaciens) used for genetically engineering plants; it is used as a vector to introduce foreign DNA into plant cells. They created the first genetically modified DNA organism. Efficient DNA sequencing methods invented by Allan Maxam no picture and Walter Gilbert (1) (1976)and by Frederick Sanger(2) (1977, developed chain termination DNA sequencing allowing scientists to read the nucleotide sequence of a DNA molecule) and his colleagues dramatically facilitated analysis of cloned DNA, and, together with the invention of the PCR by Kary Mullis (3)(1983, invented the polymerase chain reaction which is a technique enabling scientists to reproduce bits of DNA faster than ever before), information that DNA sequencing yielded about the structure and function of cloned genes led to the birth of the field of genomics. 12 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) 1996 The birth of the first cloned animal, Dolly the sheep, was announced. She was cloned by Ian Wilmut(upper), Keith Campbell(lower) and colleagues at the Roslin Institute, part of the University of Edinburgh, and the biotechnology company PPL Therapeutics near Edinburgh in Scotland, the United Kingdom. Then in the following years, Transgenic animals such as mouse, pig, cattle, lamb and transgenic plants such as tobacco, tomato, sunflower, corn, potato, soybeans were developed. However, although this has been retrospective, in reality, the accelerated scientific journey that has resulted from the ability to clone DNA has only begun. -Stanley N. Cohen Note: Genomics - a discipline in genetics that applies recombinant DNA, DNA sequencing methods, and bioinformatics to sequence, assemble, and analyze the function and structure of genomes. Genomes - the complete set of DNA within a single cell of an organism. Transgenic-containing a gene or genes transferred from another species. C. Cloning and GMO: A Comparison Cloning: Definition and Its Role Cloning is the creation of an organism that is an exact genetic copy of another. This means that every single bit of DNA is the same between the two. At its most basic level, Cloning is reproduction without sex. Sex does not refer to the act of intercourse but to sexual reproduction – the joining of genetic material from two parents into an embryo that may, if development goes well, give rise to a new adult organism. In cloning, offspring are 13 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) genetically identical to their single parent. Such offspring are the products of asexual reproduction. Cloning matters because it is on the verge of affecting daily life around the world and its importance will only grow with time. Animal cloning will revolutionize food production in the coming years and may, by turning animals into biological factories, revolutionize pharmaceutical production as well. Moving from animals to humans, cloning technology may, if some expectations prove true, radically alter medicine, leading the way to an era of personalized transplant therapies. Finally, in the longer term, it opens the door to the cloning (and potential genetic engineering) of humans, perhaps changing the very essence of what it means to be a human being. Cloning also matters because, given the field s current trajectory, it is part of our shared future. From the food supply to the medicine cabinet, cloning technology is poised to change the way we live. But these changes are controversial. Each of us can and should participate in the debates that will shape the role cloning plays in the future. Before you say yuck to drinking milk from cloned cows or rush off to save your dog s DNA in preparation for eventual cloning, take the time to learn a bit about the science. Although cloning is fairly simple, misinformation is prevalent. Understanding the science behind cloning will help make these debates more meaningful and their outcomes more satisfactory for everyone. Genetically Modified Organisms (GMO) A genetically modified organism (GMO) is the term commonly used for crops that have been genetically engineered (GE) or genetically modified (GM) to produce some desired trait. Genetic modification altered the genes to render the plant resistant either insects or herbicides. It involves the insertion into an animal of genes from another species or extra genes from the same species. GM is different from traditional breeding, where the organism's genes are manipulated indirectly; GM uses the techniques of molecular cloning and transformation to alter the structure and characteristics of genes directly. The primary focus of the research on genetic modification involves locating genes that can produce the desired results-such as those conferring insect resistance, reducing sensitivity to herbicides, increasing the amount of desired nutrients, or preventing fruits from rotting as quickly as usual. This difficult process is becoming easier with technologies that permit rapid gene sequencing and with sophisticated computer programs that can match up genetic patterns with their protein products. Molecular biologists have discovered many enzymes which change the structure of DNA in living organisms. Some of these enzymes can cut and join strands of DNA. Using such enzymes, scientists learned to cut specific genes from DNA and to build customized DNA 14 Coming Soon ONE ACTION. ONE MILLION CONSEQUENCES. (Cloning and GMOs) using these genes. They also learned about vectors, strands of DNA such as viruses, which can infect a cell and insert themselves into its DNA. With this knowledge, scientists started to build vectors which incorporated genes of their choosing and used the new vectors to insert these genes into the DNA of living organisms. Genetic engineers believe they can improve the foods we eat by doing this. For example, tomatoes are sensitive to frost. This shortens their growing season. Fish, on the other hand, survive in very cold water. Scientists identified a particular gene which enables a flounder to resist cold and used the technology of genetic engineering to insert this 'anti-freeze' gene into a tomato. This makes it possible to extend the growing season of the tomato. D. Cloning and Genetic Modification: Mechanisms Cloning The most commonly used procedure is somatic cell nuclear transfer (SCNT). This involves collecting a cell from the animal that is to be cloned (the donor cell) and removing an egg cell from another animal. This cell is enucleated, i.e. its genetic material is removed. The donor cell and the egg cell are then fused by an electrical pulse from this a cloned embryo is developed. This is implanted into a surrogate mother. In sheep and pigs, the transfer of the embryo into the surrogate mother is performed by a surgical procedure. There are a couple of ways to do cloning: artificial embryo twinning and somatic cell nuclear transfer. How do these processes differ? 1. Artificial Embryo Twinning Artificial embryo twinning is the relatively low-tech version of cloning. As the name suggests, this technology mimics the natural process of creating identical twins. Open large version In nature, twins occur just after fertilization of an egg cell by a sperm cell. In rare cases, when the resulting fertilized egg, called a zygote, tries to divide into a two-celled embryo, the two cells separate. Each cell continues dividing on its own, ultimately developing into a 15 Coming Soon ONE ACTION. ONE MILLION CONSEQUENCES. (Cloning and GMOs) separate individual within the mother. Since the two cells came from the same zygote, the resulting individuals are genetically identical. Figure 2. Two Ways of Cloning Artificial embryo twinning uses the same approach, but it occurs in a Petri dish instead of in the mother's body. This is accomplished by manually separating a very early embryo into individual cells, and then allowing each cell to divide and develop on its own. The resulting embryos are placed into a surrogate mother, where they are carried to term and delivered. Again, since all the embryos came from the same zygote, they are genetically identical. 2. Somatic Cell Nuclear Transfer Somatic cell nuclear transfer, (SCNT) uses a different approach than artificial embryo twinning, but it produces the same result: an exact clone, or genetic copy, of an individual. This was the method used to create Dolly the Sheep. What does SCNT mean? Let's take it apart: 16 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) Somatic cell: A somatic cell is any cell in the body other than the two types of reproductive cells, sperm and egg. These are also called germ cells. In mammals, every somatic cell has two complete sets of chromosomes, whereas the germ cells only have one complete set. Nuclear: The nucleus is like the cell's brain. It's an enclosed compartment that contains all the information that cells need to form an organism. This information comes in the form of DNA. It's the differences in our DNA that make each of us unique. Transfer: Moving an object from one place to another. To make Dolly, researchers isolated a somatic cell from an adult female sheep. Next, they transferred the nucleus from that cell to an egg cell from which the nucleus had been removed. After a couple of chemical tweaks, the egg cell, with its new nucleus, was behaving just like a freshly fertilized zygote. It developed into an embryo, which was implanted into a surrogate mother and carried to term. The lamb, Dolly, was an exact genetic replica of the adult female sheep that donated the somatic cell nucleus to the egg. She was the first-ever mammal to be cloned from an adult somatic cell. How does SCNT differ from the natural way of making an embryo? Open large version The fertilization of an egg by a sperm and the SCNT cloning method both result in the same thing: a dividing ball of cells, called an embryo. 17 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) Genetic Modification Figure 3. How To Build A Better Plant. Scientific American, September 2013 The most common form of genetic engineering involves the insertion of new genetic material at an unspecified location in the host genome. This is accomplished by isolating and copying the genetic material of interest using molecular cloning methods to generate a DNA sequence containing the required genetic elements for expression, and then inserting this construct into the host organism. Other forms of genetic engineering include gene targeting and knocking out specific genes via engineered nucleases such as zinc finger nucleases or engineered homing endonucleases. 18 Coming Soon ONE ACTION. ONE MILLION CONSEQUENCES. (Cloning and GMOs) Figure 4. Steps of Genetic Modification The gene for the desired trait or characteristics are identified, cut from its source and multiplied. The gene is inserted in appropriate vector to form the gene construct. A vector is like a vehicle or a carrier and has the necessary regulatory elements such as the promoter and terminator which can make the gene work. The promoter will then tell when and where the gene will be expressed and how many copies of the protein will be produced. The terminator will command the end of expression or reading of the gene. A selection gene marker is also usually in the gene construct. This will help in determining which of the bombarded tissues have incorporated the gene construct in their DNA. The gene construct is delivered to the plant cell by either of two methods. One is by coating the gene on tungsten or gold particles and delivering these particles into plant tissues by using a particle bombardment device. This device is attached to a gas tank containing helium gas which forces the DNA-coated particles into the tissues with pressure. The other method involves inserting the gene construct into Agrobacterium tumefaciens which is then used to infect a plant and eventually transfer the gene construct and its othr genes to 19 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) the plant genome. The Agrobacterium is common found in nature. If you find galls or swellings on plants; most probably, this is due to the infection of the plant with Agrobacterium. The next step is to determine which among the plant cells have integrated the introduced gene. The selection marker will do this. For example, if the selection marker is an antibiotic resistance gene marker, cells which have this gene will be able to survive in a medium containing such antibiotic. These cells are termed transformed or transgenic. Another type of selection marker gene is the green fluorescent protein or GFP marker; cells that integrate this in their DNA will produce this protein which gives off green fluorescence when beamed under a UV light. The transformed or transgenic plant tissues are allowed to grow and regenerate to complete plants. The breeder will now screen the resulting plants for the desired trait and evaluate as well their agronomic or horticultural traits. The breeder will select lines which have the desired traits and are stable. The molecular biologist/biochemist will determine the presence of the inserted gene and other biochemical characteristics of the transgenic plants. E. Products of Cloning and GMOs Products of Cloning Cloning Dolly the sheep Dolly the sheep, as the first mammal to be cloned from an adult cell, is by far the world's most famous clone. However, cloning has existed in nature since the dawn of life. From asexual bacteria to virgin birth in aphids, clones are all around us and are fundamentally no different to other organisms. A clone has the same DNA sequence as its parent and so they are genetically identical. Several clones had been produced in the lab before Dolly, including frogs, mice, and cows, which had all been cloned from the DNA from embryos. Dolly was remarkable in being the first mammal to be cloned from an adult cell. This was a major scientific achievement as it demonstrated that the DNA from adult cells, despite having specialized as one particular type of cell, can be used to create an entire organism. Dolly the cloned sheep © The Roslin institute 20 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) How Dolly was cloned Animal cloning from an adult cell is much more difficult than from an embryonic cell. So when scientists working at the Roslin Institute in Scotland produced Dolly, the only lamb born from 277 attempts, it was a major news story around the world. To produce Dolly, scientists used an udder cell from a six-year-old Finn Dorset white sheep. They had to find a way to 'reprogram' the udder cells - to keep them alive but stop them growing – which they achieved by altering the growth medium the soup in which the cells were kept alive). Then they injected the cell into an unfertilised egg cell which had had its nucleus removed, and made the cells fuse by using electrical pulses. The unfertilised egg cell came from a Scottish Blackface ewe. When the research team had managed to fuse the nucleus from the adult white sheep cell with the egg cell from the black-faced sheep, they needed to make sure that the resulting cell would develop into an embryo. They cultured it for six or seven days to see if it divided and developed normally, before implanting it into a surrogate mother, another Scottish Blackface ewe. Dolly had a white face. From 277 cell fusions, 29 early embryos developed and were implanted into 13 surrogate mothers. But only one pregnancy went to full term, and the 6.6 kg Finn Dorset lamb 6LLS (alias Dolly) was born after 148 days. What happened to Dolly? Dolly lived a pampered existence at the Roslin Institute. She mated and produced normal offspring in the normal way, showing that such cloned animals can reproduce. Born on 5 July 1996, she was euthanized on 14 February 2003, aged six and a half. Sheep can live to age 11 or 12, but Dolly suffered from arthritis in a hind leg joint and from sheep pulmonary adenomatosis, a virus-induced lung tumor that is common among sheep which are raised indoors. The DNA in the nucleus is wrapped up into chromosomes, which shorten each time the cell replicates. This meant that Dolly and her lamb, Bonnie © The Roslin institute Dolly s chromosomes were a little shorter than those of other sheep her age and her early ageing may reflect that she was raised from the nucleus of a 6-year old sheep. Dolly was also not entirely identical to her genetic mother because the mitochondria, the power plants of the cell that are kept outside the nucleus, were inherited from Dolly s egg donor mother. 21 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) Why clone sheep? Dolly the sheep was produced at the Roslin Institute as part of research into producing medicines in the milk of farm animals. Researchers have managed to transfer human genes that produce useful proteins into sheep and cows, so that they can produce, for instance, the blood clotting agent factor IX to treat haemophilia or alpha-1-antitrypsin to treat cystic fibrosis and other lung conditions. Inserting these genes into animals is a difficult and laborious process; cloning allows researchers to only do this once and clone the resulting transgenic animal to build up a breeding stock. The development of cloning technology has led to new ways to produce medicines and is improving our understanding of development and genetics. Since Dolly Since 1996, when Dolly was born, other sheep have been cloned from adult cells, as have cats, rabbits, horses and donkeys, pigs, goats and cattle. In 2004 a mouse was cloned using a nucleus from an olfactory neuron, showing that the donor nucleus can come from a tissue of the body that does not normally divide. Improvements in the technique have meant that the cloning of animals is becoming cheaper and more reliable. This has created a market for commercial services offering to clone pets or elite breeding livestock, but still with a $100,000 price-tag. The advances made through cloning animals have led to a potential new therapy to prevent mitochondrial diseases in humans being passed from mother to child. About 1 in 6000 people is born with faulty mitochondria, which can result in diseases like muscular dystrophy. To prevent this, genetic material from the embryo is extracted and placed in an egg cell donated by another woman, which contains functioning mitochondria. This is the same process as used in cloning of embryonic cells of animals. Without this intervention, the faulty mitochondria are certain to pass on to the next generation. The treatment is currently not permitted for use in humans. However, the Human Fertilisation & Embryology Authority in the UK has reported that there is general support in the public for legalising the therapy and making it available to patients. Why was Dolly Created? The development of the cloning technology was an extension of Roslin Institute's interest in the application of transgenic technology to farm animals. Transgenic mice have been available since early 1980s produced by a very sophisticated method of genetic modification through a technology using embryonic stem cells. Cells in culture can be genetically modified in very precise ways: removing genes, substituting one 22 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) gene for another, introducing a single base pair change in the genetic code. In mice it was possible to genetically modify these cells, introduce them into a mouse embryo and the resulting mice that are born would be chimeric with some normal cells, some genetically modified cells. At least some of the offspring of these chimeras would contain the very precise genetic modification. Since embryonic stem cells had not been isolated from farm animals, this method of genetic modification was not available. Cloning was therefore a potential alternative way of achieving the same end. Why was Roslin Institute interested in genetically modifying farm animals? Since mid-1980s there has been a research interest in developing new uses for farm animals and one of those research ideas being pursued since the early days was the idea of producing human proteins in the milk of transgenic cattle or sheep. Those experiments used a very simple technique for genetic modification called pronuclear injection. This involved introducing the DNA construct, the human gene coding for the protein of interest, into a recently fertilised egg and taking that early embryo to term. A very small proportion of animals produced in this way carried the gene and a proportion of this small proportion expressed the gene so that human protein was produced in the milk. This was a very inefficient way of genetic modification. There was no control over where gene was inserted or indeed how many genes were inserted and it was only possible to add genes. As part of the developing interest in this area there was a need to improve the efficiency of genetic modification, to control gene expression more reliably and ensure it was expressed in particular tissues only. Why was this research done at Roslin Institute? People in the past have been motivated to try cloning as a means of replicating the very best animals with respect to agricultural production. Can you copy the very best bulls? And that was the motivation behind the work that Steen Willardsen had done in Texas in the 1980s. In Roslin Institute's case the motivation, at least initially, to pursue nuclear transfer was a very practical application in terms of developing a new way of genetically modifying animals. You might expect this work to have been in mice, and there is a history of this work being done in amphibians but it wasn't successful. Ultimately the interest in this area of science waned, the lack of technical success re-enforced the view that differentiated cells were not reprogrammable and it was only our particular interest in a rather narrow practical field that maintained our commitment to the area. If the research community that uses mice had taken an interest, there's probably hundreds of labs around the world that could have cloned mice but there are only six or seven research institutes around the world that have experience in embryo transfer, IVF technology, the sort of understanding of cattle or sheep reproduction that was a basic requirement for Roslin's success. 23 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) Why was Dolly Important? The birth of Dolly overturned the assumption among scientists that the whole process of differentiation was irreversible. We all start life as a single cell, the fertilised egg. The cell divides and multiplies and by the time we are born, there are maybe 200 different cell types, each containing the same DNA, the same 30,000 or so genes, but each has differentiated into a particular role. That role is determined by the proportion of active genes within the cell that determines whether the cell is for example a liver cell or a nerve cell. A presumption among cell biologists was that this was a one way process of progressive and permanent change. What Dolly demonstrated was that it is possible to take a differentiated cell and essentially turn its clock back; to reactivate all its silent genes and make the cell behave as though it was a recently fertilised egg. Dolly was also important because she captured the public imagination. A clone, a copy has been a very discernible strand within science fiction. The idea that there might be and exact copy of oneself somewhere around is a theme that has been pursued in science fiction and the prospect that it might be possible to clone a human being excited a lot of speculation and interest. What is the longterm significance of Dolly? At the moment that's difficult to say. The practical applications of cloning, of copying livestock seem relatively limited. The likelihood is that the longer lasting benefit will be in the change in perception about biology. Our understanding now is that the cells in our bodies are a lot more plastic than we previously thought and it may be that as we understand more about repair processes, for various organs and tissues, we might find that this understanding informs research that is able to augment the bodies normal repair mechanisms. It may well prove to be an important factor in stem cell research and allow the derivation of stem cells from tissues other than early human embryos. This would alleviate the reservations that many people have about the use of human embryos for research or therapeutic purposes. Noah the Gaur A rare Asian Ox called a Gaur was successfully cloned Sioux Center Iowa in 2003. It was successfully cloned and gestated in the womb of a cow named Bessie which is a scientific first. The project was particularly interesting as it coupled cloning with that of interspecies birth. The researchers hope that technique may be able to be used to shore up animal population. 24 Coming Soon ONE ACTION. ONE MILLION CONSEQUENCES. (Cloning and GMOs) The steps involved in this process are as follows: 1. Remove DNA from a unfertilized cow egg. 2. Insert full DNA strand from Gaur into the empty egg. 3. Apply small electrical pulses to fuse. 4. Add chemicals to induce fertilisation events. 5. Place fertilized back inside cow s uterus. This process was repeated five times but only Noah made it to the late stages of fetal Development the other four were unfortunately spontaneously aborted. After Ten months of hard work by the scientists Noah the Gaur was born in Iowa. Unfortunately he died after 48 hours of life from Dysentry, "We don't think it had to do with the cloning, 'Dysentery affects farm animals" Robert Lanza Vice president of scientific development at the center said. A Family of Pigs: Millie, Alexis, Christa, Dotcom, and Carrel Blacksburg, VA PPL Therapeutics Inc is pleased to announce that on 5th March 2000, five piglets, all healthy, were born as a result of nuclear transfer (cloning) using adult cells. This is the first time cloned pigs have been successfully produced from adult cells. DNA from blood samples taken from the piglets was shown in independent tests to be identical to DNA from the cells used to produce the piglets but clearly different from DNA taken from the surrogate mother. The DNA tests were carried out by Celera-AgGEN on coded samples. The cell samples had been provided to the testing company before the piglets were born. 25 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) The successful cloning of these pigs is a major step in achieving PPL s xenograft objectives. It opens the door to making modified pigs whose organs and cells can be successfully transplanted into humans; the only near term solution to solving the worldwide organ shortage crisis. Pigs are the preferred species for xenotransplantation on scientific and ethical grounds. Clinical trials could start in as little as four years and analysts believe the market could be worth $6 billion for solid organs alone, with as much again possible from cellular therapies, eg. transplantable cells that produce insulin for treatment of diabetes. Nuclear transfer in pigs has proved to be more difficult than for other livestock, in part because pig reproductive biology is inherently more intractable, and partly because pigs need a minimum number of viable fetuses to maintain pregnancy, whereas sheep and cows, for example, need only one. The method used to produce the five female piglets, to be named Millie, Christa, Alexis, Carrel and Dotcom, was different from that used to produce "Dolly" in that it used additional inventive steps for which a patent application has been filed. The work was carried out by PPL s US staff in Blacksburg, Virginia, partly supported by an ATP Award from the US Government s National Institute of Standards and Technology. This award has as its objective the production of a "knock-out" pig, i.e. a pig which has a specific gene inactivated. The ability to clone pigs is the first essential step in achieving this objective. The gene to be inactivated is alpha 1-3 gal transferase. This gene is responsible for adding to pig cells a particular sugar group recognized by the human immune system as foreign and which therefore triggers an immune response leading to hyperacute rejection in humans of the transplanted organ. PPL has already achieved the required targeted gene knock out in pig cells. Tetra the Rhesus Monkey Tetra is neither the first monkey clone, nor the first mammal to be cloned by embryo splitting. 26 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) Tetra was produced by a technique called "embryo splitting." Here's how it works:    An egg from a mother and sperm from a father are used to create a fertilized egg. After the embryo grows into eight cells, researchers split it into four identical embryos, each consisting of just two cells. The four embryos are then implanted into surrogate mothers. Schatten said that in effect, a single embryo becomes four embryos, all genetically identical. Nonetheless, the birth of this animal does suggest a new and possibly easier and cheaper method of cloning non-human primates. This accomplishment could prove to be a boon to medical researchers and could be a step towards human cloning. Tetra is the name given to the one monkey that survived of four identical embryos that were implanted in four separate host mothers. Using a procedure similar to that used in in vitro fertilization,.scientists at the Oregon Regional Primate Research Center began by taking an egg from the mother monkey and sperm from the father monkey and then mixing them together to create a fertilized egg. Once the embryo had grown into eight cells, the scientists then divided the embryo into four identical embryos consisting of two cells each. These four embryos were then implanted into four potential monkey mothers. Tetra, from the Greek word for four, was the result. This is not the first time this technique has been used to create mammalian twins. The same technique is already being used in cattle. A physician also reported using the technique to clone human embryos as far back as 1993. Nor it is the first time that monkeys have been cloned. Researchers from the same Oregon research group rerpoted cloning a monkey in 1997 using the nuclear transfer method. That method involves removing a set of chromosomes from each of the eight cells in a primitive monkey embryo and then inserting into egg cells from which the original DNA had been removed. These embryos were then implanted in the wombs of host mothers using in vitro fertilization techniques. What is new about the creation of Tetra is that this is the first time researchers have created a perfect genetic copy of a monkey by using the embryo splitting technique. Unlike the earlier monkey clone, Tetra is the first to possess both identical nuclear and cytoplasmic components. This offers researchers for the first time the opportunity to produce a line of identical primates for medical research. This would allow them to test new treatments for a variety of conditions such as AIDS, cancer and heart disease in a way that is not currently possible. The birth of Tetra suggests that scientists may have bridged the scientific gap between genetically identical knockout mice and human patients. There are many potential areas where this technology might advance biological research. In addition to providing 27 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) researchers with more reliable test animals and controls, the discovery could also benefit researchers looking at the role of the maternal environment in the characteristics of offspring. It is also likely to be a boon to those studying embryology and stem cell development. Many questions remain to be answered. For example, researchers still need to confirm that twins or 'multiples' created by this method are as health and long-lived as normal monkeys. They also need to explore why the success rate has been so low. It is also worth noting that, although laboratory tests did show that Tetra was identical to the embryos that did not survive, this is one step short from producing two living identical clones from separate mothers. The research is ongoing. Four pregnancies, each with a viable fetus, have been established from the last seven embryo transfers of identical twins.. One pregnancy is from the transfer of a single embryo, the other three are singletons resulting from the transfer of two unrelated embryos. If successful, these identical twins will be named Romulus and Rhesus. The research appeared in the Jan. 14, 2000 issue of Science. Products of Genetic Modification Genetically Engineered Cow Produces World's First Hypoallergenic Milk The calf completely lacked a milk protein called betalactoglobulin. It also lacked a tail. Cow genes could be modified to prevent the animals from producing proteins that cause allergic reactions, according to a new study. Scientists in New Zealand engineered a 28 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) dairy cow to lack the milk protein beta-lactoglobulin, while other milk proteins were dramatically increased. The team used RNA interference to inhibit the expression of certain genes that code for the production of BLG, which causes allergic reactions in people and isn't found in human milk. They tested it on mice first, and then engineered a cow egg cell's nucleus to express the same micro RNAs that shut down BLG. This engineered ovum was fertilized and implanted into a surrogate mother. The team started with 57 embryos and ultimately got one healthy calf, but unexpectedly, it was born with no tail. The researchers believe this mutation is unrelated to the transgenic change, but they still need to figure out exactly what caused it. Finally, the team gave the calf hormones to make it produce milk early, and they found the milk contained no BLG. The work shows that RNA interference could be an effective way to modify livestock to have desirable traits, the researchers say. Meanwhile, the researchers are waiting until the calf gets a little older to study the mystery of its missing tail. Their paper is published in the Proceedings of the National Academy of Sciences. ANDi, Genetically Modified Monkey Oregon researchers have created the first genetically modified monkey. ANDi, a playful, coffee-colored rhesus monkey born on October 2nd 2000, has been engineered to carry a gene from another species. OSHU named the monkey ANDi because it stands for inserted DNA spelled backward. ANDi was born with an extra glowing gene called Green Fluorescent Protein (GFP). This GFP gene, which is naturally occurring in jellyfish, was taken from a jellyfish and genetically added to ANDi s DNA sequence through 29 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) his chromosomes. OSHU used Rhesus monkeys because they share 95% of the same genes as humans. The work demonstrates that a foreign gene can be delivered and inserted into a primate chromosome. The researchers anticipate that gene insertions in the monkey will lead to primate models of human diseases—like Alzheimer's, diabetes, heart disease and obesity— that will offer a more robust testing ground for new drugs, gene therapy and modified stem cells. To create ANDi, Chan and his colleagues injected 224 unfertilized rhesus eggs with a virus carrying the green fluorescent protein (GFP) gene. The virus's job is to integrate the gene into a random site on one of the chromosomes. Six hours later, each egg was artificially fertilized by sperm injection. Roughly half of the fertilized eggs grew and divided, reaching the four-cell stage. Forty were chosen and implanted into twenty surrogate mothers—two per mother. Of these, three healthy males were born and two twin males were stillborn. ANDi was the only live monkey carrying the GFP gene. Cloning and GMO Advantages and Disadvantages Medical Advantages of Cloning Although there are many potential downsides, and many people feel uncertain about whether or not this practice is morally right, the advantages of cloning are numerous. Certain types of cloning may be used to create food sources with a higher nutritional value, while others may be used to create types of medicine or treatments. One of the bestknown advantages of cloning is organ transplantation, which could potentially save the lives of accident victims and of those waiting for an organ donation. The medical advantages of cloning may begin with the actual nourishment of the body. Not only can cloned cows and chickens produce more eggs and milk, but scientists may also be able to genetically alter the nutritional value of these foods. Infants incapable of breastfeeding may also benefit from certain types of animal cloning. For instance, a cow whose genetic code is manipulated may produce milk that contains proteins similar to those found in human breast milk. Fertility is another one of the possible advantages of cloning. For those who are sterile, this solution may provide hope where other options have failed. One of the most common processes of reproductive cloning begins by injecting the genetic material of one parent into an egg. Once this is done, the egg is stimulated by electricity or chemicals, and then placed into the uterus. Although this process has been accomplished to some degree in animals, further research is needed to see whether or not it will also work for humans. 30 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) Besides nutrition and fertility, the advantages of cloning reach into treating and possibly curing many medical issues. Organ transplantation is the best-known medical use for cloning. Sometimes transplanted organs are harvested from animals, which are regularly rejected by human recipients. Cloned animals, however, may bear human genes, which may make rejection less common. Cloning may also be beneficial in replacing bone, cartilage, and skin in burn and accident victims. Other medical advantages of cloning consist of the creation of advanced medicine. These medicines may be used for heart and bone marrow treatments, to control diabetes and rheumatoid arthritis, and perhaps even to cure kidney conditions and Parkinson's disease. In addition, cloning might also be able to cure certain types of cancer by replacing mutated genes with healthy, normal ones. This process often consists of taking immune cells from the patient's own body, duplicating them, and then placing them back into the system. Advantages 1. Potential benefits to modern medicine Given the fact that the cells can be manipulated to mimic other types of cells, this can provide new ways to treat diseases like cancer and Alzheimer s. Cloning also offers hope to persons needing organ transplants. People requiring organ transplants to survive an illness often wait years for a suitable donor. In many cases these patients die waiting, as there are long lists of people requiring organs. Theoretically, cloning could eliminate this by producing more animals that can act as suitable donors. 2. Helping infertile couples Cloning offers couples dealing with fertility the chance to have a child of their own. Many infertile couples can t be helped by the techniques currently available. In fact, although some states have already banned human cloning because of ethical issues, more couples struggling to have children are starting to consider the possibilities that cloning offer. 3. Reverse the aging process Cloning is being touted as a future answer to reverse the effects of aging. The anti-aging market is a prime target because it is already a multibillion industry. 4. Protecting Endangered Species Despite the best efforts of conservationists worldwide, some species are nearing extinction. The successful cloning of Dolly represents the first step in protecting endangered wildlife. 31 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) 5. Improving food supply Cloning could provide a means of cultivating plants that are stronger and more resistant to diseases, while producing more. The same could happen to livestock as well where diseases such as foot and mouth disease could be eradicated. Cloning could therefore effectively solve the world s food problem and minimize or possible eradicate starvation. Disadvantages of Cloning Cloning is defined as using the cells of one living subject, plant or animal, to create another duplicate subject. A cloned subject will be identical to its parent. Cloning has become the center of a huge debate over the advantages and disadvantages of producing clones, especially of animals and humans. While this technology could be useful for laboratory studies and for creating desirable livestock, there are several disadvantages of cloning that should be considered. One of the biggest disadvantages of cloning is that the technology is still so uncertain. Dolly the sheep, the first mammalian clone, was born in 1996. While she was initially successful, she died young of a disease not normally seen in sheep of her age. Scientists are still unsure of any genetic mutations that might occur when an animal is cloned. Also, while Dolly was a successful clone, there were hundreds of failed clones before she was made, including several dead fetuses. Other cloned animals have turned out horribly deformed. Losing gene diversity is another of the disadvantages of cloning. Gene diversity is what keeps an entire species from being wiped out by a singular virus if none of them have natural immunities. This is due to the lack of gene diversity. Gene mutations happen naturally, and help to explain why some people naturally are taller, shorter, or more athletic than others. Some people and animals naturally have a stronger immune system. If gene diversity is lost due to excessive cloning, there are no mutations to allow some of the cloned group to survive a newly introduced disease. Another of the disadvantages of cloning is that there are a lot of ethical considerations that would cause most people to protest. One of these ethical concerns is that cloning is unnatural, and considered playing God. Another concern is the treatment of clones. Clones would have the same needs as non-clones of their species. Humane treatment guidelines would still apply. There is always a risk of cloning technology being abused. One of the main disadvantages of cloning is that the technology would have to be kept closely monitored. For example, imagine what a corrupt dictator could do with cloning. There will always be someone looking to use cloning for their own personal use, and many feel that the best way to prevent this is to not pursue cloning at all. 32 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) There are many advantages to cloning, such as the chance of curing certain diseases and being able to breed ideal stock for research and consumption. However, the disadvantages of cloning are seen by many to far outweigh any benefits that might be seen. Because of the risk taking involved in cloning, it is a technology that many experts say may be better left alone, at least until it is better understood. Disadvantages 1. The Element of Uncertainty While the cloning of Dolly was seen as a success story, many embryos were destroyed before the desired result was achieved. The process started with 277 eggs, and Dolly was the single successful outcome. Regardless of success in other areas, the field of cloning still has a long way to go. Infertile couples for example, could go through the same heartache as they would if in vitro fertilization failed. 2. Inheriting diseases Cloning creates a copy of the original. A human clone would therefore inherit the genetic traits of its predecessor. This includes genetic abnormalities and diseases. Dolly the sheep for example exhibited signs of what some suggested were premature aging, although this was firmly denied by her developers . 3. The Potential for Abuse If human cloning became a reality what checks and balances would be put in place to prevent abuse? Would scientists go overboard with the technology? If a couple has a clone that they are not happy with, what would they do next? These are all questions that must be raised in any discussion on cloning. Some have expressed the view that clones could be grown in a farm-like fashion simply for harvesting organs or stem cells. The potential for devaluing human life cannot be ignored. Advantages of GMOs The mapping of genetic material for GMO crops increased knowledge of genetic alterations and introduced the ability to enhance genes in crops to make them more advantageous for human consumption and production. For example, plants can be engineered to be temperature resistant or produce higher yields. This provides greater genetic diversity in different regions where climate limits productivity. Another good reason to have GMO crops planted is to add nutritional value to crops that lack necessary vitamins and nutrients. There are areas around the world that rely on rice or corn crops, and other plant genes may be added to the crop to increase the 33 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) nutritional value of that food. This will help malnourished populations receive more nutrients from their diet. We have already made pesticide resistant plants so that farmers can use the right kinds of pesticides to rid insects and not inhibit plant growth. This will increase crop yield in two ways; there will be fewer insects and pests to eat the crops, and they will grow without being bothered by pesticides. Advantages Crops • Enhanced taste and quality • Increased nutrients, yields, and stress tolerance • New products and growing techniques • Reduced maturation time • Improved resistance to disease, pests, and herbicides Animals • Better yields of meat, eggs, and milk • Increased resistance, productivity, hardiness, and feed efficiency • "Friendly" bioherbicides and bioinsecticides • Bioprocessing for forestry products • More efficient processing • Improved animal health and diagnostic methods Environment • Conservation of soil, water, and energy • Better natural waste management • Genes can also be manipulated in trees to absorb more CO2 and reduce the threat of global warming. Society • Increased food security for growing populations 34 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) Disadvantages of GMOs The GMO process includes adding new genetic material into an organism's genome. In agricultural ecology, similar to bacterial genetic engineering, this means introducing new genes in the genome of crops like corn. Experimental plantings of GMO crops began in Canada and the U.S. in the s. The first time it became large scale commercial cultivation was in the mid s. Research on the effects of large scale cultivation of GM crops sparked various concerns. These ideas are brought up in different research studies conducted on ecosystems with GMO strains. GMO strains have the potential to change our agriculture. A plant with unwanted or residual effects that might remain in the soil for extended period of time. European Union agricultural regulators were alerted by Morrissey s research that GM strains from GM crops remained in the soil for years after the crop was removed. Data reported that despite the absence of the GM plant, the strain persisted for up to six years. Engineered plants can act as mediators to transfer genes to wild plants and then create weeds. To keep these new weeds under control scientists invented new GMO weed herbicides that were not necessary for non GMO weeds. These chemicals are toxic to various amphibians and mammals, such as cows feeding on GMO crops. In vivo tests show that the uptake of herbicides has toxic consequences on certain organisms. There is opposition in the introduction of GM genes on genetic diversity. The GM genes from crops can spread to organic farm crops and threaten crop diversity in agriculture. If crop diversity decreases, this affects the entire ecosystem and impacts the population dynamics of other organisms. The chance that one genetically modified crop strain could pollinate an already existent non-GM crop is unlikely and unpredictable. There are many conditions that must be met for cross pollination to occur. However, when a large scale plantation releases a GM strain during pollination, this risk increases. The cross pollination to non-GM plants could create a hybrid strain, which means there is a greater possibility of ecological novelty, or new artificial strains being introduced into the environment that could potentially reduce biodiversity through competition. Disadvantages Food regulatory authorities require that GM foods receive individual pre-market safety assessments. Also, the principle of substantial equivalence is used. This means that an existing food is compared with its genetically modified counterpart to find any differences between the existing food and the new product. The assessment investigates: • Toxicity (using similar methods to those used for conventional foods). 35 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) • Tendency to provoke any allergic reaction. • Whether there is any nutritional deficit or change in the GM food. • Stability of the inserted gene. • Any other unintended effects of the gene insertion. F. The Future of Cloning and GMO: Researches and Recommendations Cloning and GMO: Researches The study entitled Production of a Calf from a Nuclear Transfer Embryo Using in Vitro Matured Oocytes written by Ushijima, H., et. al. was conducted to examine the possibility of nuclear transplantation using the bovine oocytes matured in vitro. Sixteen-cell stage embryos were assigned to donor cells. The follicular oocytes matured in vitro were used as enucleated recipient oocytes. The enucleated oocytes were fused with a blastomere from donor embryos by electrofusion. Most recipient oocytes fused with donor cells. The reconstituted eggs developed to 2-cell stage when cultured on a layer of cumulus cells (187, 58%). Out of the 325 reconstituted eggs, 35 (11%) developed into morulae and 9 (3%) into blastocysts stage respectively. Eighteen morula or blastocyst stage embryos were transferred nonsurgically to 9 recipient cows into 1-3 embryos per recipient. Two recipients were confirmed pregnant. One of recipients produced a live offspring resulting from a fresh donor blastomere. The study showed that in vitro matured oocytes can be used as recipient cytoplasm. The study entitled Successful Mouse Cloning of an Outbred Strain by Trichostatin A Treatment after Somatic Nuclear Transfer written by Kishigami, S. et. al. was conducted to test the validity of trichostatin A (TSA) cloning technique in which the researchers tried to clone the adult ICR mouse, an outbred strain, which has never been directly cloned before. The researchers obtained both male and female cloned mice from cumulus and fibroblast cells of adult ICR mice with4-5% percent success rates when TSA was used, which is comparable to 5-7% of B6D2F1. Thus, the TSA treatment was the first cloning technique to allow the researchers to successfully clone outbred mice, demonstrating that this technique not only improves the success of cloning from hybrid strains, but also enables mouse cloning from normally uncloned strains. The study entitled Cloned cows with short telomeres deliver healthy offspring with normal-length telomeres written by Miyashita, N. et. al. was conducted to investigate longetivity and lifetime performance in cloned animals. The researchers produced cloned cows with short telomeres using oviductal ephitelial cells as donor cells. At 5 years of age, despite despite the prescence of short telomeres, all cloned cows deliver multiple healthy offspring following artificial insemination with conventionally processed spermatozoa 36 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) from noncloned bulls, and their milk production was comparable to that of donor cows. Moreover, the study revealed that the offspring had normal-length telomeres in their leukocytes and major organs. Thus, cloned animals have normal function germ lines, and therefore germ line function can completely restore telomere lengths in clone gametes by telomerase activity, resulting in healthy offspring with normal-length telomeres. The study entitled Application of Genetically Modified and Cloned Pigs in Translational Research written by Matsunari, H. et. al. reviews the current status and future prospects of genetically modified and cloned pigs in translational research. It also highlights pig especially designed as disease models, for xenotransplantation or to carry cell marker genes. Finally, use of porcine somatic stem and progenitor cells in preclinical studies of cell transplantation study therapy is also discussed. The study entitled Mouse Cloning Using a Drop of Peripheral Blood written by Kamimura, S. et. al. was conducted to determine whether peripheral blood cells freshly collected from living mice could be used for SCNT. The researchers collected a drop of peripheral blood (15– μl from the tail of a donor. A nucleated cell leukocyte suspension was prepared by lysing the red blood cells. Following SCNT using randomly selected leukocyte nuclei, cloned offspring were born at a 2.8% birth rate. Fluorescenceactivated cell sorting revealed that granulocytes/monocytes and lymphocytes could be roughly distinguished by their sizes, the former being significantly larger. The researchers then cloned putative granulocytes/monocytes and lymphocytes separately, and obtained 2.1% and 1.7% birth rates, respectively (P > 0.05). Because the use of lymphocyte nuclei inevitably results in the birth of offspring with DNA rearrangements, the researchers applied granulocyte/monocyte cloning to two genetically modified strains and two recombinant inbred strains. Normal-looking offspring were obtained from all four strains tested. The present study clearly indicated that genetic copies of mice could be produced using a drop of peripheral blood from living donors. This strategy will be applied to the rescue of infertile founder animals or a last-of-line animal possessing invaluable genetic resources. Future of Cloning and GMO: Conclusion and Recommendations Cloning is a break through technology that improves the variety and breed of plants and animals. It is important because it is on the verge of affecting daily life around the world and its importance will only grow with time. Animal cloning will revolutionize food production in the coming years and may, by turning animals into biological factories, revolutionize pharmaceutical production as well. Moving from animals to humans, cloning technology may, if some expectations prove true, radically alter medicine, leading the way to an era of personalized transplant therapies. Finally, in the longer term, it opens the door to the cloning and potential genetic engineering of humans, perhaps changing the very 37 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) essence of what it means to be a human being. When Dolly was cloned in 1996, the research was primarily funded by a biotechnology firm that aimed to revolutionize the way drugs are produced. The basic idea is to create, through cloning, genetically modified sheep or cows that produce therapeutic compounds, such as insulin or growth hormone, in their milk. Pharmaceutical companies could isolate these valuable compounds from the milk for a fraction of the cost of traditional manufacturing methods. The milk would not be intended for human consumption and would probably be discarded after the therapeutics had been isolated. This technique, known as pharming, offers potential economic benefits for drug companies and has taken off since Dolly s birth. Numerous cows have been bred to produce therapeutics in their milk and some scientists are exploring the possibility of harvesting drugs from other body fluids, including urine. Pharming raises a number of concerns, including the risk of drugproducing animals accidentally entering the food supply. Although the risks may be remote, even those of us unfazed by drinking milk from a cloned cow wouldn t be pleased to find out the milk was significantly enriched with a prescription medicine. While cloned animals that produce therapeutic compounds already exist, the creation of cloned human embryos to facilitate medical therapies remains in the future and raises serious ethical questions. Many scientists are optimistic that cloning will, one day, regularly be used to create stem cells genetically matched to specific patients. These cells could, potentially, help treat a range of debilitating conditions, such as type 1 diabetes and Parkinson s disease. Because the cells would be genetically matched to the individual patient, they might avoid the immune rejection problems that complicate transplant therapies today. This potential therapeutic technique is controversial, however, because deriving these patient-matched stem cells, using currently envisioned approaches, would require the creation of a cloned human embryo. At five days of age, the stem cells would be isolated from the embryo and the developmental process halted. Dramatic advances toward this vision of regenerative medicine were reported by a group of researchers based in South Korea, but in late 2005 the veracity of this work was called into question: today, it is clear that most, if not all, these advances were fraudulent. Despite this set-back, many scientists believe the vision remains promising and therapeutic cloning is being pursued by scientists around the world. Genetic modification has become a routine part of biotechnology, and it is being increasingly relied upon. In certain areas, such as producing drugs and food-modifying enzymes, the potential for serious problems to arise seems small, but when used in food crops, there are some evident dangers from the fact that these crops become so widespread in the world environment. The level of alarm felt by some people about risks from eating these food crops is most likely exaggerated. However, as the technology develops further, attention must be paid to possible rare adverse responses to foods, especially allergenic 38 ONE ACTION. ONE MILLION CONSEQUENCES. Coming Soon (Cloning and GMOs) responses, before commercial production begins. Cloning also matters because, given the field s current trajectory, it is part of our shared future. From the food supply to the medicine cabinet, cloning technology is poised to change the way we live. But these changes are controversial. Each of us can and should participate or conduct researchers that will shape the role cloning plays in the future. Cloning, like so many other issues that have faced modern science, must be carefully evaluated. There will always be detractors, those who feel that anyone involved in cloning is playing God. And this may not be too far from the truth. However, any discussion on cloning must be looked at in the context of its inherent value to mankind. The choice is ours. We cannot ignore gene technology, nor should we condemn all of it. The key is proper regulation. Either we control gene technology today, or gene technology will redesign us by tomorrow. 39