Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/134040
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Type: | Journal article |
Title: | The Nestin neural enhancer is essential for normal levels of endogenous Nestin in neuroprogenitors but is not required for embryo development |
Author: | Thomson, E. Dawson, R. H'ng, C.H. Adikusuma, F. Piltz, S. Thomas, P.Q. |
Citation: | PLoS One, 2021; 16(11):e0258538-1-e0258538-16 |
Publisher: | Public Library of Science |
Issue Date: | 2021 |
ISSN: | 1932-6203 1932-6203 |
Editor: | Varshney, G. |
Statement of Responsibility: | Ella Thomson, Ruby Dawson, Chee Ho H'ng, Fatwa Adikusuma, Sandra Piltz, Paul Q. Thomas |
Abstract: | 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. |
Keywords: | Nestin ; Gene expression; Homozygosity; Central nervous system; CRISPRs; Embryos; Immunohistochemistry techniques; Polymerase chain reaction |
Rights: | © 2021 Thomson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
DOI: | 10.1371/journal.pone.0258538 |
Grant ID: | http://purl.org/au-research/grants/nhmrc/DP140103965 |
Published version: | http://dx.doi.org/10.1371/journal.pone.0258538 |
Appears in Collections: | Genetics publications |
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hdl_134040.pdf | Published version | 2.43 MB | Adobe PDF | View/Open |
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