Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Fukuda, Kei, Okuda, Akihiko, Yusa, Kosuke, Shinkai, Yoichi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2018
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
_version_ 1783329841981947904
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
work_keys_str_mv AT fukudakei acrisprknockoutscreenidentifiessetdb1targetretroelementsilencingfactorsinembryonicstemcells
AT okudaakihiko acrisprknockoutscreenidentifiessetdb1targetretroelementsilencingfactorsinembryonicstemcells
AT yusakosuke acrisprknockoutscreenidentifiessetdb1targetretroelementsilencingfactorsinembryonicstemcells
AT shinkaiyoichi acrisprknockoutscreenidentifiessetdb1targetretroelementsilencingfactorsinembryonicstemcells
AT fukudakei crisprknockoutscreenidentifiessetdb1targetretroelementsilencingfactorsinembryonicstemcells
AT okudaakihiko crisprknockoutscreenidentifiessetdb1targetretroelementsilencingfactorsinembryonicstemcells
AT yusakosuke crisprknockoutscreenidentifiessetdb1targetretroelementsilencingfactorsinembryonicstemcells
AT shinkaiyoichi crisprknockoutscreenidentifiessetdb1targetretroelementsilencingfactorsinembryonicstemcells