The Nestin neural enhancer is essential for normal levels of endogenous Nestin in neuroprogenitors but is not required for embryo development
| العنوان: | The Nestin neural enhancer is essential for normal levels of endogenous Nestin in neuroprogenitors but is not required for embryo development |
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| المؤلفون: | Ruby Dawson, Ella Thomson, Paul Q. Thomas, Chee Ho H’ng, Fatwa Adikusuma, Sandra Piltz |
| المصدر: | PLoS ONE, Vol 16, Iss 11, p e0258538 (2021) PLoS ONE PLoS ONE, Vol 16, Iss 11 (2021) |
| بيانات النشر: | Public Library of Science (PLoS), 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Central Nervous System, Embryology, Heredity, Gene Expression, Artificial Gene Amplification and Extension, Biochemistry, Nervous System, Synthetic Genome Editing, Polymerase Chain Reaction, Homozygosity, Genome Engineering, Nestin, Mice, 0302 clinical medicine, Pregnancy, Gene expression, Medicine and Health Sciences, Frameshift Mutation, Neurons, 0303 health sciences, Multidisciplinary, 030302 biochemistry & molecular biology, Crispr, Gene Expression Regulation, Developmental, Neural stem cell, Cell biology, Enhancer Elements, Genetic, Essential gene, Engineering and Technology, Medicine, Female, Synthetic Biology, Anatomy, Research Article, Science, Embryonic Development, Repressor, Mice, Transgenic, Bioengineering, Biology, Research and Analysis Methods, 03 medical and health sciences, Genetics, Animals, Molecular Biology Techniques, Enhancer, Molecular Biology, Gene, 030304 developmental biology, Embryos, Biology and Life Sciences, Proteins, Synthetic Genomics, Embryonic stem cell, Introns, Cytoskeletal Proteins, Mutation, 030217 neurology & neurosurgery, Developmental Biology |
| الوصف: | Enhancers are vitally important during embryonic development to control the spatial and temporal expression of genes. Recently, large scale genome projects have identified a vast number of putative developmental regulatory elements. However, the proportion of these that have been functionally assessed is relatively low. While enhancers have traditionally been studied using reporter assays, this approach does not characterise their contribution to endogenous gene expression. We have studied the murine Nestin (Nes) intron 2 enhancer, which is widely used to direct exogenous gene expression within neural progenitor cells in cultured cells and in vivo. We generated CRISPR deletions of the enhancer region in mice and assessed their impact on Nes expression during embryonic development. Loss of the Nes neural enhancer significantly reduced Nes expression in the developing CNS by as much as 82%. By assessing NES protein localization, we also show that this enhancer region contains repressor element(s) that inhibit Nes expression within the vasculature. Previous reports have stated that Nes is an essential gene, and its loss causes embryonic lethality. We also generated 2 independent Nes null lines and show that both develop without any obvious phenotypic effects. Finally, through crossing of null and enhancer deletion mice we provide evidence of trans-chromosomal interaction of the Nes enhancer and promoter. |
| اللغة: | English |
| تدمد: | 1932-6203 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::dfa8d2e2245eaab85f3ddb17460f0624 https://doaj.org/article/9cf4d4f009da49f8b62d4a8421930bbe |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....dfa8d2e2245eaab85f3ddb17460f0624 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19326203 |
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