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21 pages, 1921 KiB  
Article
Enhancing Solanum lycopersicum Resilience: Bacterial Cellulose Alleviates Low Irrigation Stress and Boosts Nutrient Uptake
by Noelia De la Cruz Gómez, César Poza-Carrión, Lucía Del Castillo-González, Ángel Isidro Martínez Sánchez, Ana Moliner, Inmaculada Aranaz and Marta Berrocal-Lobo
Plants 2024, 13(15), 2158; https://doi.org/10.3390/plants13152158 - 4 Aug 2024
Viewed by 1024
Abstract
The use of natural-origin biomaterials in bioengineering has led to innovative approaches in agroforestry. Bacterial cellulose (BC), sharing the same chemical formula as plant-origin cellulose (PC), exhibits significantly different biochemical properties, including a high degree of crystallinity and superior water retention capacity. Previous [...] Read more.
The use of natural-origin biomaterials in bioengineering has led to innovative approaches in agroforestry. Bacterial cellulose (BC), sharing the same chemical formula as plant-origin cellulose (PC), exhibits significantly different biochemical properties, including a high degree of crystallinity and superior water retention capacity. Previous research showed that natural-origin glucose-based chitin enhanced plant growth in both herbaceous and non-herbaceous plants. In this study, we produced BC in the laboratory and investigated its effects on the substrate and on Solanum lycopersicum seedlings. Soil amended with BC increased root growth compared with untreated seedlings. Additionally, under limited irrigation conditions, BC increased global developmental parameters including fresh and dry weight, as well as total carbon and nitrogen content. Under non-irrigation conditions, BC contributed substantially to plant survival. RNA sequencing (Illumina®) on BC-treated seedlings revealed that BC, despite its bacterial origin, did not stress the plants, confirming its innocuous nature, and it lightly induced genes related to root development and cell division as well as inhibition of stress responses and defense. The presence of BC in the organic substrate increased soil availability of phosphorus (P), iron (Fe), and potassium (K), correlating with enhanced nutrient uptake in plants. Our results demonstrate the potential of BC for improving soil nutrient availability and plant tolerance to low irrigation, making it valuable for agricultural and forestry purposes in the context of global warming. Full article
(This article belongs to the Topic Plant Responses to Environmental Stress)
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Figure 1
<p>ATR-FTIR spectrum of bacterial cellulose (BC). (<b>A</b>) Whole spectrum. (<b>B</b>) Detail of the spectra between 650 and 1000 cm<sup>−1</sup> showing two bacterial cellulose bands around 750 and around 710 cm<sup>−1</sup>, corresponding to the presence of the crystalline phases Iα and Iβ of BC, respectively. The presence of stars in FTIR spectra denotes the beta linkage of cellulose monomers. (<b>C</b>) XRD diffractogram recorded for the isolated bacterial cellulose, showing the crystalline structure of cellulose I with three main peaks hightlighted with stars located at 14.7, 17.3, and 23.2 corresponding to crystallographic planes 100, 010, and 110, respectively.</p>
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<p>Physiological effects of bacterial cellulose (BC) on <span class="html-italic">Solanum lycopersicum</span> (Sl). (<b>A</b>) Seed germination rate on substrate containing 0.01% (<span class="html-italic">w</span>:<span class="html-italic">w</span>), BC (triangle), or watered control (circle). Data were collected up to 14 days. (<b>B</b>) Seed germination rate under hydroponic conditions in the presence of 0.01% (<span class="html-italic">w</span>:<span class="html-italic">v</span>), BC (triangle), or watered control (circle). Data were collected up to 14 days. (<b>C</b>) Total fresh and dry weight (g) of seedlings, growth under optimal irrigation conditions (I), or under regulated non-irrigation conditions (NI), in the absence (white) or presence of BC (grey). (<b>D</b>) Representative photos of 14-day-old seedlings growing at the same plate under I or NI conditions, in the absence (−) or presence (+) of BC. Bars: 1 cm. (<b>E</b>) Total shoot (left figure) and root (right figure) lengths (cm), measured in the absence (white) or presence of BC (grey) growth under I or NI conditions. (<b>F</b>) Detail of shoots and roots photos corresponding to (<b>E</b>), obtained for measuring seedling growth using ImageJ<sup>®</sup> tool (1.53 version) (see <a href="#sec4-plants-13-02158" class="html-sec">Section 4</a>) under I or NI conditions. Assays were performed at least three times with similar results using ten seedlings per pot and twelve pots per plate and tree plates per treatment (n = 120 per plate). Data were analyzed with the Stat-graphics Centurion 19 program, using a Variance check (<span class="html-italic">p</span> &gt; 0.05) and a non-parametric Kruskal–Wallis test. * Significant statistical differences.</p>
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<p>Effect of bacterial cellulose (BC) on Fe, K, and P content in plants and substrate. (<b>A</b>) Effect of BC (0.01%, <span class="html-italic">w</span>:<span class="html-italic">w</span>) on total P content on substrate (S, mg/kg), plants (P, g/100 g), or planted substrate (PS, mg/kg) in the absence (C) or presence of BC (0.01%, <span class="html-italic">w</span>:<span class="html-italic">w</span>). (<b>B</b>) Effect of BC on total Fe content on S (mg/kg), P (mg/Kg × 10<sup>2</sup>), or PS (mg/kg) in the absence (C) or presence of BC. (<b>C</b>) Effect of BC on total K content on S (mg/kg), P (g/100 g), or PS (mg/kg) in the absence (C) or presence of BC. Substrate and seedling samples were collected after fourteen days, making the treatment at time zero. At least three assays were performed analyzing ten seedlings per pot, three pots per treatment, with three plates (n = 120). Data were analyzed with the Stat-graphics Centurion 19 program, using a Variance check (<span class="html-italic">p</span> &gt; 0.05) and a non-parametric Kruskal–Wallis’s test. White is control (C), while the grey bars correspond to bacterial cellulose (BC) treatment. * Significant statistical differences.</p>
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<p>RNA sequencing data analysis of <span class="html-italic">Solanum lycopersicum</span> seedlings responding to bacterial cellulose (BC). (<b>A</b>) Volcano scatter plots obtained with BioJupies<sup>®</sup> software, displaying the log2-fold changes calculated by performing a differential gene expression analysis (see <a href="#sec4-plants-13-02158" class="html-sec">Section 4</a>). Red points indicate significantly up-regulated genes, while blue points indicate down-regulated genes. The upper plot shows results of seedlings responding to BC (0.01%, <span class="html-italic">w</span>:<span class="html-italic">v</span>) after one hour, related to watered non-treated seedlings. The lower plot shows a positive control for molecular plant response to chitin (Qq, 0.01%, <span class="html-italic">w</span>:<span class="html-italic">v</span>) (see <a href="#sec4-plants-13-02158" class="html-sec">Section 4</a>). (<b>B</b>) Heatmap obtained with BioJupies<sup>®</sup> software (see <a href="#sec4-plants-13-02158" class="html-sec">Section 4</a>), displaying significant gene expression on selected genes in the RNA-seq dataset. Induced (red) or repressed genes (blue) (FDR&lt; or &gt;0.05) are shown after 1 h of treatment with BC. Every row of the heatmap represents a gene, and every column represents triplicates of controls (C1 to C3) and BC treatments (BC1 to BC3), correspondingly. Every cell displays normalized gene expression values. <span class="html-italic">Solanum lycopersicum</span> gene IDs are shown in the left column obtained by using the ITAG 3.2 genome annotation version. Black dots mean significant diferences on BC treated plants related to untreated plants lower than 1.5.</p>
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16 pages, 3777 KiB  
Article
Taxonomic Identification of the Arctic Strain Nocardioides Arcticus Sp. Nov. and Global Transcriptomic Analysis in Response to Hydrogen Peroxide Stress
by Bailin Cong, Hui Zhang, Shuang Li, Shenghao Liu, Jing Lin, Aifang Deng, Wenqi Liu and Yan Yang
Int. J. Mol. Sci. 2023, 24(18), 13943; https://doi.org/10.3390/ijms241813943 - 11 Sep 2023
Viewed by 1341
Abstract
Microorganisms living in polar regions rely on specialized mechanisms to adapt to extreme environments. The study of their stress adaptation mechanisms is a hot topic in international microbiology research. In this study, a bacterial strain (Arc9.136) isolated from Arctic marine sediments was selected [...] Read more.
Microorganisms living in polar regions rely on specialized mechanisms to adapt to extreme environments. The study of their stress adaptation mechanisms is a hot topic in international microbiology research. In this study, a bacterial strain (Arc9.136) isolated from Arctic marine sediments was selected to implement polyphasic taxonomic identification based on factors such as genetic characteristics, physiological and biochemical properties, and chemical composition. The results showed that strain Arc9.136 is classified to the genus Nocardioides, for which the name Nocardioides arcticus sp. nov. is proposed. The ozone hole over the Arctic leads to increased ultraviolet (UV-B) radiation, and low temperatures lead to increased dissolved content in seawater. These extreme environmental conditions result in oxidative stress, inducing a strong response in microorganisms. Based on the functional classification of significantly differentially expressed genes under 1 mM H2O2 stress, we suspect that Arc9.136 may respond to oxidative stress through the following strategies: (1) efficient utilization of various carbon sources to improve carbohydrate transport and metabolism; (2) altering ion transport and metabolism by decreasing the uptake of divalent iron (to avoid the Fenton reaction) and increasing the utilization of trivalent iron (to maintain intracellular iron homeostasis); (3) increasing the level of cell replication, DNA repair, and defense functions, repairing DNA damage caused by H2O2; (4) and changing the composition of lipids in the cell membrane and reducing the sensitivity of lipid peroxidation. This study provides insights into the stress resistance mechanisms of microorganisms in extreme environments and highlights the potential for developing low-temperature active microbial resources. Full article
(This article belongs to the Section Molecular Microbiology)
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<p>Phylogenetic relationship of strain Arc9.136. Neighbor-joining phylogenetic tree showing the position of the isolate Arc9.136 within the genus <span class="html-italic">Nocardioides</span> based on 16S rDNA gene sequence data. Bootstrap ≥ 1000 (accession number: OP861529.2).</p>
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<p>Whole-genome assembly and annotation of Arc9.136. (<b>A</b>) Circular map of the Arc9.136 genome. From inside to outside, the first innermost circle represents genome GC skew values, the second circle describes genome GC content, the third circle shows ncRNA, the fourth, fifth, and sixth circles are GO, KEGG, and COG function annotations, respectively. The seventh circle is genes. (<b>B</b>) GO analysis of Arc9.136.</p>
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<p>Transcriptome sequencing of Arc9.136 under 1 mM H<sub>2</sub>O<sub>2</sub>. (<b>A</b>) Cluster analysis of the H<sub>2</sub>O<sub>2</sub>-treated group (group B: H-1, H-2, H-3) and nontreated group (group A: 1-1, 1-2, 1-3). The length of the branch represents the distance between samples. (<b>B</b>) Volcano plot showing the DEGs between the H<sub>2</sub>O<sub>2</sub>-treated group (group B: H-1, H-2, H-3) and nontreated group (group A: 1-1, 1-2, 1-3). The X-axis indicates the fold change in gene expression (threshold, |log2 (treatment/control)| ≥ 0.5), while the Y-axis indicates the statistically significant level (threshold, <span class="html-italic">p</span> value ≤ 0.05). (<b>C</b>) Function enrichment scatter plot of DEGs by COG annotation. The rich factor represents the ratio of the number of target genes to the total number of annotated genes in this pathway. (<b>D</b>) Histogram analysis of DEGs by COG annotation.</p>
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<p>Verification of gene expression levels by real-time qRT-PCR analysis. Significant difference indicated by ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Scanning electron microscopy (SEM) images of Arc9.136. Scale (<b>A</b>) 200 μm; Scale (<b>B</b>) 500 nm.</p>
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36 pages, 11448 KiB  
Article
Proteo-Trancriptomic Analyses Reveal a Large Expansion of Metalloprotease-Like Proteins in Atypical Venom Vesicles of the Wasp Meteorus pulchricornis (Braconidae)
by Jean-Luc Gatti, Maya Belghazi, Fabrice Legeai, Marc Ravallec, Marie Frayssinet, Stéphanie Robin, Djibril Aboubakar-Souna, Ramasamy Srinivasan, Manuele Tamò, Marylène Poirié and Anne-Nathalie Volkoff
Toxins 2021, 13(7), 502; https://doi.org/10.3390/toxins13070502 - 19 Jul 2021
Cited by 6 | Viewed by 3951
Abstract
Meteorus pulchricornis (Ichneumonoidea, Braconidae) is an endoparasitoid wasp of lepidopteran caterpillars. Its parasitic success relies on vesicles (named M. pulchricornis Virus-Like Particles or MpVLPs) that are synthesized in the venom gland and injected into the parasitoid host along with the venom during oviposition. [...] Read more.
Meteorus pulchricornis (Ichneumonoidea, Braconidae) is an endoparasitoid wasp of lepidopteran caterpillars. Its parasitic success relies on vesicles (named M. pulchricornis Virus-Like Particles or MpVLPs) that are synthesized in the venom gland and injected into the parasitoid host along with the venom during oviposition. In order to define the content and understand the biogenesis of these atypical vesicles, we performed a transcriptome analysis of the venom gland and a proteomic analysis of the venom and purified MpVLPs. About half of the MpVLPs and soluble venom proteins identified were unknown and no similarity with any known viral sequence was found. However, MpVLPs contained a large number of proteins labelled as metalloproteinases while the most abundant protein family in the soluble venom was that of proteins containing the Domain of Unknown Function DUF-4803. The high number of these proteins identified suggests that a large expansion of these two protein families occurred in M. pulchricornis. Therefore, although the exact mechanism of MpVLPs formation remains to be elucidated, these vesicles appear to be “metalloproteinase bombs” that may have several physiological roles in the host including modifying the functions of its immune cells. The role of DUF4803 proteins, also present in the venom of other braconids, remains to be clarified. Full article
(This article belongs to the Special Issue Evolution, Genomics and Proteomics of Venom)
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<p>The venom apparatus of <span class="html-italic">Meteorus pulchricornis</span>. From the female wasp (<b>A</b>) the venom apparatus (<b>B</b>) was obtained by pulling out the ovipositor (ovi; white arrow in A). The ovarioles (ov), the two filamentous venom glands (<span class="html-italic">vgf</span>) and the large unique milky venom reservoir (<span class="html-italic">r</span>) are clearly visible. (<b>C</b>) TEM observation of a cross section of a venom gland filament showing a glandular cell with the secretory cell glandular canal (<span class="html-italic">cgc</span>) at its center that ends in the lumen of the collecting gland duct (<span class="html-italic">lu</span>). The nucleus (<span class="html-italic">nu</span>) surrounded by its nuclear envelope had and apparent normal shape. (<b>D</b>) MpVLPs may derive from the small electron dense vesicles contained within large cytoplasmic vesicles (<span class="html-italic">Lv</span>) found associated with the Golgi apparatus (<span class="html-italic">Gg</span>). (<b>E</b>) The internal canal (<span class="html-italic">cgc</span>) of the secretory gland cell, lined by microvilli, is filled with secreted mature MpVLPs. (<b>F</b>) MpVLPs purified by centrifugation of venom collected from <span class="html-italic">M. pulchricornis</span> venom reservoir (MpVLPs average longer diameter 250 nm, <span class="html-italic">n</span> = 20). These vesicles were apparently single membraned vesicles (membrane mean width 30 ± 13 nm, <span class="html-italic">n</span> = 17).</p>
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<p>Species classified by the number of hits reported for <span class="html-italic">M. pulchricornis</span> CDS protein sequences against the NCBI NR database.</p>
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<p>Protein profile of total venom (TV) compared to the supernatant (Super) and the MpVLPs pellet fractions obtained after centrifugation. A total of 25 and 13 bands were cut off for TV and the washed 15,000 g Pellets (MpVLPs) lanes, respectively, to be submitted to the MS-MS analysis. Left and right lanes correspond to molecular weight markers (MW in kDa). 12% SDS-PAGE in reducing conditions and gel was silver stained.</p>
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<p>Sequence alignment of nucleotidases from <span class="html-italic">M. pulchricornis</span>, human, <span class="html-italic">E.coli</span>, snake, drosophila and bee. Important conserved residues or motifs are shaded in grey or boxed. Functionally important residues are marked below the alignment with the following labels: “R” indicates the three arginine in the active site of the <span class="html-italic">E. coli</span> enzyme; “F”, the phenylalanines that bind the adenine moiety; “S” the residues that interact with the substrate and “s” those involved in non-polar interactions (from [<a href="#B61-toxins-13-00502" class="html-bibr">61</a>]). The only two cysteines in MpNUC that can form a bond are in red. USHA.ECOLI, Protein UshA <span class="html-italic">Escherichia coli</span> P07024 (uniport); 5NTD.HUMAN, 5’-nucleotidase <span class="html-italic">Homo sapiens</span> P21589; V5NTD.CROAD, <span class="html-italic">Crotalus adamanteus</span> venom 5’-nucleotidase F8S0Z7; NT5E-2PA, <span class="html-italic">D. melanogaster</span> Q8SZY4; 5NUC.Apis, protein 5NUC <span class="html-italic">Apis mellifera</span> XP.394018 (NCBI). Identical amino acids are indicated by a star, conservation by a colon and substitution by a dot.</p>
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<p>Alignment of the putative <span class="html-italic">M. pulchricornis</span> venom lipocalins. Grey boxes indicate the three putative lipocalin structural regions (SCR1, 2, 3). In green, the hydrophobic tryptophan (W) retrieved in SCRs, in red the cysteines. Identical amino acids are indicated by a star, conservation by a colon and substitution by a dot.</p>
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<p>Clustal alignment of the <span class="html-italic">M. pulchricornis</span> GGTs with the human (CAG30380.1) and <span class="html-italic">A. ervi</span> (CAL69624.1) GGT1 sequences. The catalytic threonine residue that forms the N-terminus of the small subunit (T381) after autocleavage to form the mature heterodimeric enzyme is highlighted with a black box containing a white “T” with an arrow above. Residues proposed to interact with glutathione in human GGT1 are shaded in grey and residues that significantly reduced human GGT enzymatic activity by site-directed mutagenesis are in red. Identical amino acids are indicated by a star, conservation by a colon and substitution by a dot.</p>
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<p>Sequence Alignments for Pancreatic Lipase Sequences. Key active site residues from Human Pancreatic lipase-related protein 2 (P54317) are in red shaded in grey; in shaded grey, amino acids conserved around the active site residues. The underline human sequence residues represent the two bonds between cysteines. Identical amino acids are indicated by a star, conservation by a colon and substitution by a dot.</p>
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<p>Alignment of hyaluronidases from <span class="html-italic">M. pulchricornis</span> (Hyal-Mp from contig 293), Human (Hyaluronidase 2, HYAL2-HUMAN, Q12891), bee (<span class="html-italic">Apis mellifera</span>, Q08169) and <span class="html-italic">Cotesia congregata</span> (CCQ71107). The putative active-site residues Asp(D), Glu(E), and Glu(E) (shaded in grey) of the hyaluronidase were conserved in the <span class="html-italic">M. pulchricornis</span> sequence, although a conservative replacement E → N occurred that may affect its activity [<a href="#B81-toxins-13-00502" class="html-bibr">81</a>]. The four cysteine residues forming two disulfide bridges are also conserved (in red). Identical amino acids are indicated by a star, conservation by a colon and substitution by a dot.</p>
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<p>Alignment of the first CAP domain of contig 2455 (Mp-CRISP) with selected sequences of hymenoptera antigen 5 allergens. The CAP signature motifs are shaded in grey and conserved residues that form the putative active site are shaded in cyan [<a href="#B85-toxins-13-00502" class="html-bibr">85</a>]. Cysteine residues that form disulphide bridges in Ag5 proteins are marked in red. Asterisks, colons and periods indicate identical, conserved and semi-conserved residues, respectively. Sequence from <span class="html-italic">Vespula vulgaris</span> (VA5.VESVU, Q05110.1), <span class="html-italic">Dolichovespula maculata</span> (bald-faced hornet)(VA52.DOLMA, P10736.1), <span class="html-italic">Polistes dominula</span> (European paper wasp) (VA5.polistes, NP.001310265.1), and <span class="html-italic">Microctonus hyperodae</span> (A9YME1.1).</p>
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<p>Amino acid sequence alignments of contig 590 protein (Mp_SERPIN) and human SERPINA1 (P01009|A1AT_HUMAN Alpha-1-antitrypsin). The RCL position of the human sequence is underlined. The cleavage site is situated between the position P1 (Met) and P1′ (Ser) of the human sequence, only the Ser is conserved in Mp_SERPIN. The important Alanine-rich hinge site shaded in grey is also absent from contig 590. Asterisks, colons and periods indicate identical, conserved and semi-conserved residues, respectively.</p>
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13 pages, 521 KiB  
Review
Approaching Shared Pathophysiology in Immune-Mediated Diseases through Functional Genomics
by David González-Serna, Gonzalo Villanueva-Martin, Marialbert Acosta-Herrera, Ana Márquez and Javier Martín
Genes 2020, 11(12), 1482; https://doi.org/10.3390/genes11121482 - 9 Dec 2020
Cited by 8 | Viewed by 2736
Abstract
Immune-mediated diseases (IMDs) are complex pathologies that are strongly influenced by environmental and genetic factors. Associations between genetic loci and susceptibility to these diseases have been widely studied, and hundreds of risk variants have emerged during the last two decades, with researchers observing [...] Read more.
Immune-mediated diseases (IMDs) are complex pathologies that are strongly influenced by environmental and genetic factors. Associations between genetic loci and susceptibility to these diseases have been widely studied, and hundreds of risk variants have emerged during the last two decades, with researchers observing a shared genetic pattern among them. Nevertheless, the pathological mechanism behind these associations remains a challenge that has just started to be understood thanks to functional genomic approaches. Transcriptomics, regulatory elements, chromatin interactome, as well as the experimental characterization of genomic findings, constitute key elements in the emerging understandings of how genetics affects the etiopathogenesis of IMDs. In this review, we will focus on the latest advances in the field of functional genomics, centering our attention on systemic rheumatic IMDs. Full article
(This article belongs to the Section Molecular Genetics and Genomics)
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<p>Overview of different techniques used in functional genomics. These techniques cover the spectrum from early association studies, through techniques that explore the physical interaction of these variants and their effects on the transcriptome, to phenotype characterization and gene function. GWAS: genome-wide association studies; WES: whole-exome sequencing; WGS: whole-genome sequencing; sc-RNA-seq: single-cell RNA sequencing, TALEs: Transcription activator-like effectors; CRISPR: clustered regularly interspaced short palindromic repeats; Cas9: CRISPR associated protein 9.</p>
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20 pages, 2270 KiB  
Review
Transcriptomics in Alzheimer’s Disease: Aspects and Challenges
by Eva Bagyinszky, Vo Van Giau and SeongSoo A. An
Int. J. Mol. Sci. 2020, 21(10), 3517; https://doi.org/10.3390/ijms21103517 - 15 May 2020
Cited by 46 | Viewed by 8242
Abstract
Alzheimer’s disease (AD) is the most common cause of dementia. Although the heritability of AD is high, the knowledge of the disease-associated genes, their expression, and their disease-related pathways remain limited. Hence, finding the association between gene dysfunctions and pathological mechanisms, such as [...] Read more.
Alzheimer’s disease (AD) is the most common cause of dementia. Although the heritability of AD is high, the knowledge of the disease-associated genes, their expression, and their disease-related pathways remain limited. Hence, finding the association between gene dysfunctions and pathological mechanisms, such as neuronal transports, APP processing, calcium homeostasis, and impairment in mitochondria, should be crucial. Emerging studies have revealed that changes in gene expression and gene regulation may have a strong impact on neurodegeneration. The mRNA–transcription factor interactions, non-coding RNAs, alternative splicing, or copy number variants could also play a role in disease onset. These facts suggest that understanding the impact of transcriptomes in AD may improve the disease diagnosis and also the therapies. In this review, we highlight recent transcriptome investigations in multifactorial AD, with emphasis on the insights emerging at their interface. Full article
(This article belongs to the Special Issue Molecular Genetic of Alzheimer's Disease)
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<p>Overview showing the workflow for transcriptomic studying in Alzheimer’s disease, from transcriptomic data generation to integration of regulatory information to assess gene regulatory networks.</p>
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<p>Methodological aspects to consider in most common miRNA biomarkers for Alzheimer’s research.</p>
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