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A CRISPR knockout screen identifies SETDB1-target retroelement silencing factors in embryonic stem cells
In mouse embryonic stem cells (mESCs), the expression of provirus and endogenous retroelements is epigenetically repressed. Although many cellular factors involved in retroelement silencing have been identified, the complete molecular mechanism remains elusive. In this study, we performed a genome-w...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5991520/ https://www.ncbi.nlm.nih.gov/pubmed/29728365 http://dx.doi.org/10.1101/gr.227280.117 |
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author | Fukuda, Kei Okuda, Akihiko Yusa, Kosuke Shinkai, Yoichi |
author_facet | Fukuda, Kei Okuda, Akihiko Yusa, Kosuke Shinkai, Yoichi |
author_sort | Fukuda, Kei |
collection | PubMed |
description | In mouse embryonic stem cells (mESCs), the expression of provirus and endogenous retroelements is epigenetically repressed. Although many cellular factors involved in retroelement silencing have been identified, the complete molecular mechanism remains elusive. In this study, we performed a genome-wide CRISPR screen to advance our understanding of retroelement silencing in mESCs. The Moloney murine leukemia virus (MLV)–based retroviral vector MSCV-GFP, which is repressed by the SETDB1/TRIM28 pathway in mESCs, was used as a reporter provirus, and we identified more than 80 genes involved in this process. In particular, ATF7IP and the BAF complex components are linked with the repression of most of the SETDB1 targets. We characterized two factors, MORC2A and RESF1, of which RESF1 is a novel molecule in retroelement silencing. Although both factors are recruited to repress provirus, their roles in repression are different. MORC2A appears to function dependent on repressive epigenetic modifications, while RESF1 regulates repressive epigenetic modifications associated with SETDB1. Our genome-wide CRISPR screen cataloged genes which function at different levels in silencing of SETDB1-target retroelements and provides a useful resource for further molecular studies. |
format | Online Article Text |
id | pubmed-5991520 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-59915202018-12-01 A CRISPR knockout screen identifies SETDB1-target retroelement silencing factors in embryonic stem cells Fukuda, Kei Okuda, Akihiko Yusa, Kosuke Shinkai, Yoichi Genome Res Research In mouse embryonic stem cells (mESCs), the expression of provirus and endogenous retroelements is epigenetically repressed. Although many cellular factors involved in retroelement silencing have been identified, the complete molecular mechanism remains elusive. In this study, we performed a genome-wide CRISPR screen to advance our understanding of retroelement silencing in mESCs. The Moloney murine leukemia virus (MLV)–based retroviral vector MSCV-GFP, which is repressed by the SETDB1/TRIM28 pathway in mESCs, was used as a reporter provirus, and we identified more than 80 genes involved in this process. In particular, ATF7IP and the BAF complex components are linked with the repression of most of the SETDB1 targets. We characterized two factors, MORC2A and RESF1, of which RESF1 is a novel molecule in retroelement silencing. Although both factors are recruited to repress provirus, their roles in repression are different. MORC2A appears to function dependent on repressive epigenetic modifications, while RESF1 regulates repressive epigenetic modifications associated with SETDB1. Our genome-wide CRISPR screen cataloged genes which function at different levels in silencing of SETDB1-target retroelements and provides a useful resource for further molecular studies. Cold Spring Harbor Laboratory Press 2018-06 /pmc/articles/PMC5991520/ /pubmed/29728365 http://dx.doi.org/10.1101/gr.227280.117 Text en © 2018 Fukuda et al.; Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genome.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. |
spellingShingle | Research Fukuda, Kei Okuda, Akihiko Yusa, Kosuke Shinkai, Yoichi A CRISPR knockout screen identifies SETDB1-target retroelement silencing factors in embryonic stem cells |
title | A CRISPR knockout screen identifies SETDB1-target retroelement silencing factors in embryonic stem cells |
title_full | A CRISPR knockout screen identifies SETDB1-target retroelement silencing factors in embryonic stem cells |
title_fullStr | A CRISPR knockout screen identifies SETDB1-target retroelement silencing factors in embryonic stem cells |
title_full_unstemmed | A CRISPR knockout screen identifies SETDB1-target retroelement silencing factors in embryonic stem cells |
title_short | A CRISPR knockout screen identifies SETDB1-target retroelement silencing factors in embryonic stem cells |
title_sort | crispr knockout screen identifies setdb1-target retroelement silencing factors in embryonic stem cells |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5991520/ https://www.ncbi.nlm.nih.gov/pubmed/29728365 http://dx.doi.org/10.1101/gr.227280.117 |
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