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    Embryonic Stem Cells

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    Takwa Shlaki
    This document discusses embryonic stem cell-mediated gene transfer. It begins by introducing embryonic stem cells and their ability to transfer genes. It then outlines the steps of the process, which involves introducing DNA into ES cells, selecting transformed cells, injecting them into mouse embryos, and testing offspring for the desired gene. Advantages include precise gene targeting and testing at the early cell stage. Disadvantages are the difficulty and expense of maintaining ES cell lines. Finally, applications of transgenic mice produced through this method are discussed, such as for disease models and biotechnology.

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    Embryonic Stem Cells

    Uploaded by

    Takwa Shlaki
    41 views6 pages
    This document discusses embryonic stem cell-mediated gene transfer. It begins by introducing embryonic stem cells and their ability to transfer genes. It then outlines the steps of the process, which involves introducing DNA into ES cells, selecting transformed cells, injecting them into mouse embryos, and testing offspring for the desired gene. Advantages include precise gene targeting and testing at the early cell stage. Disadvantages are the difficulty and expense of maintaining ES cell lines. Finally, applications of transgenic mice produced through this method are discussed, such as for disease models and biotechnology.

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    Embryonic stem cells

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    This document discusses embryonic stem cell-mediated gene transfer. It begins by introducing embryonic stem cells and their ability to transfer genes. It then outlines the steps of the process, which involves introducing DNA into ES cells, selecting transformed cells, injecting them into mouse embryos, and testing offspring for the desired gene. Advantages include precise gene targeting and testing at the early cell stage. Disadvantages are the difficulty and expense of maintaining ES cell lines. Finally, applications of transgenic mice produced through this method are discussed, such as for disease models and biotechnology.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    41 views6 pages

    Embryonic Stem Cells

    Uploaded by

    Takwa Shlaki
    This document discusses embryonic stem cell-mediated gene transfer. It begins by introducing embryonic stem cells and their ability to transfer genes. It then outlines the steps of the process, which involves introducing DNA into ES cells, selecting transformed cells, injecting them into mouse embryos, and testing offspring for the desired gene. Advantages include precise gene targeting and testing at the early cell stage. Disadvantages are the difficulty and expense of maintaining ES cell lines. Finally, applications of transgenic mice produced through this method are discussed, such as for disease models and biotechnology.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    of 6

    ‫كلية التقنية الطبية صرمان‬

    ‫قسم التقنيات الحيوية والهندسة الوراثية الطبية‬

    ‫‪Embryonic stem cells‬‬


    ‫‪mediated gene transfer‬‬

    ‫‪By : Takwa Morad Alshlaki‬‬


    Introduction

    Embryonic stem (ES) cells mediated gene transfer:


    Embryonic stem (ES) cells mediated gene transfer method based on findings of
    Gossler (1986).
    It’s a transgenic technique have had a profound impact on genetic analysis in the
    mouse during the past decade. powerful tool to investigate the molecular regulation
    of gene expression, development, and disease.
    ES cells are pluripotent cells, found in the inner cell mass (ICM) of embryos at
    the blastocyst stage of development. These cells have not yet differentiated and
    maintain the ability to develop into any type of tissue during the embryonic and
    foetal development.
    DNA can be introduced into the ES cells in vitro . embryonic stem cells grown
    at blastocyst stage, containing the desired DNA are incorporated into the host’s
    embryo and then embryo inserted in the uterus of a surrogate mother, resulting
    in a chimeric animal.
    The method uses homologous recombination of DNA to permit precise
    targeting of DNA in embryonic stem cells.
    If the homologous sequence to be introduced into the cell carries a mutation or
    a gene from another species, the new sequence will replace the specific
    targeted gene. This is the method of choice for gene inactivation therefore
    called as "knock-out" method, particular important for the study of the genetic
    control of developmental processes.
    Steps of Embryonic stem (ES) cells mediated gene
    transfer :
    1. Using recombinant DNA methods, build molecules of DNA containing the
    structural gene you desire (e.g, the insulin gene), vector DNA to enable the
    molecules to be inserted into host DNA molecules, promoter and enhancer
    sequences to enable the gene to be expressed by host cells.

    2. Transform ES cells in culture to expose cultured cells to the DNA so that


    some will incorporate it.

    3. Select for successfully transformed cells.

    4. Inject these cells into the inner cell mass (ICM) of mouse blastocysts.

    5. Embryo transfer.

    6. Prepare a pseudopregnant The stimulus of mating elicits the hormonal


    changes needed to make her uterus receptive.

    7. Transfer the embryos into her uterus.

    8. If the implant successfully and develop into healthy pups (no more than
    one-third will), Test her offspring.
    9. Remove a small piece of tissue from the tail and examine its DNA for the
    desired gene. No more than 10- 20% will have it, and they will be
    heterozygous for the gene.
    10. Establish a transgenic strain.

    11. Bred two heterozygous mice and screen their offspring for the 1:4 that will
    be homozygous for the transgene.

    12. Mating these will found the transgenic strain.


    Advantages:
    1. Application of gene targeting thus enabling site-directed
    insertion of DNA .
    2. Gene targeting involves inducing the embryonic stem
    cell to remove one of its own genes and replace it with a
    modified version of the same gene.
    3. This method allows testing for transgenes at the early
    cell stage.
    4. Embryonic stem (ES) cells are relatively efficient in
    homologous recombination in comparison to other
    animal cells.
    5. To detect precisely mutations in the gene via
    homologous recombination

    Disadvantages:
    1. Production, characterization and maintenance of
    pluripotent Embryonic stem (ES) cells lines difficult.
    2.The procedure is time consuming
    3. expensive
    Application of Transgenic mice produced by Embryonic
    stem (ES) cells mediated gene transfer :

    1.Transgenic animals are used in medical research in as disease model.


    2.Transgenic animals are also used in the field called toxicology as a
    source of detection of toxicants.
    3.They are also used in Biotechnology as they produce specific proteins.
    4.They are also used in Xenografting.
    5.In molecular biology, they are used for analyzing regulation of gene
    expression.
    6.Transgenic animals have a great role in pharmaceutical industry,
    They are used for drug testing and drug production.

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