Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis

Fujita, Toshitsugu; Kitaura, Fusako; Oji, Asami; Tanigawa, Naoki; Yuno, Miyuki; Ikawa, Masahito; Taniuchi, Ichiro; Fujii, Hodaka

doi:https://doi.org/10.1111/gtc.12573

Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis

Fujita, Toshitsugu; Kitaura, Fusako; Oji, Asami et al.

Genes to Cells, 2018, 23(4), 318-325

Number of Access:555（2025-08-22 01:06 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/78574

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Title	Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis
Creator	NRID 1000010550030 1000010550030 CreatorName Fujita, Toshitsugu
	CreatorName Kitaura, Fusako
	NRID 1000050803417 1000050803417 CreatorName Oji, Asami
	CreatorName Tanigawa, Naoki
	CreatorName Yuno, Miyuki
	NRID 1000020304066 1000020304066 ORCID 0000-0001-9859-6217 0000-0001-9859-6217 researchmap permalink read0054910 read0054910 Scopus Author ID 16689054500 16689054500 CreatorName Ikawa, Masahito
	NRID 1000020284573 1000020284573 CreatorName Taniuchi, Ichiro
	NRID 1000030302665 1000030302665 CreatorName Fujii, Hodaka
Subject	ChIP
	chromatin immunoprecipitation
	dCas9
	enChIP
	transgenic mouse
Description	Fujita, T, Kitaura, F, Oji, A, et al. Transgenic mouse lines expressing the 3xFLAG‐dCas9 protein for enChIP analysis. Genes Cells. 2018; 23: 318– 325. https://doi.org/10.1111/gtc.12573
Abstract	We developed the engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) technology to isolate specific genomic regions while retaining their molecular interactions. In enChIP, the locus of interest is tagged with an engineered DNA-binding molecule, such as a modified form of the clustered regularly interspaced short palindromic repeats (CRISPR) system containing a guide RNA (gRNA) and a catalytically inactive form of Cas9 (dCas9). The locus is then affinity-purified to enable identification of associated molecules. In this study, we generated transgenic mice expressing 3xFLAG-tagged Streptococcus pyogenes dCas9 (3xFLAG-dCas9) and retrovirally transduced gRNA into primary CD4+ T cells from these mice for enChIP. Using this approach, we achieved high yields of enChIP at the targeted genomic region. Our novel transgenic mouse lines provide a valuable tool for enChIP analysis in primary mouse cells.
Publisher	Blackwell Publishing Ltd
Source Title	Genes to Cells
Volume (Issue)	23(4)
Page	318-325
Date of Issued	2018-04-10
Language	English
Handle URL	https://hdl.handle.net/11094/78574
PISSN	13569597
eISSN	13652443
NCID	AA11623816
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1111/gtc.12573
Relation.isIdenticalTo	PMID 29480524
Access Rights	open access
CopyRight	© 2018 The Authors. Genes to Cells published by Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd. This article is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
oaire:version	Version of Record
Category	学術雑誌論文 Journal Article

Resource Type	journal article
Local Type	Journal Article
DCTERMS.bibliographicCitation	Genes to Cells. 2018, 23(4), p. 318-325
DC.title	Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis
DC.creator	Fujita, Toshitsugu
	Kitaura, Fusako
	Oji, Asami
	Tanigawa, Naoki
	Yuno, Miyuki
	Ikawa, Masahito
	Taniuchi, Ichiro
	Fujii, Hodaka
DC.publisher	Blackwell Publishing Ltd
DC.language' scheme='DCTERMS.RFC1766	English
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2018-04-10
DC.identifier	https://doi.org/10.1111/gtc.12573
DC.subject	ChIP
	chromatin immunoprecipitation
	dCas9
	enChIP
	transgenic mouse
DC.description	Fujita, T, Kitaura, F, Oji, A, et al. Transgenic mouse lines expressing the 3xFLAG‐dCas9 protein for enChIP analysis. Genes Cells. 2018; 23: 318– 325. https://doi.org/10.1111/gtc.12573
DCTERMS.abstract	We developed the engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) technology to isolate specific genomic regions while retaining their molecular interactions. In enChIP, the locus of interest is tagged with an engineered DNA-binding molecule, such as a modified form of the clustered regularly interspaced short palindromic repeats (CRISPR) system containing a guide RNA (gRNA) and a catalytically inactive form of Cas9 (dCas9). The locus is then affinity-purified to enable identification of associated molecules. In this study, we generated transgenic mice expressing 3xFLAG-tagged Streptococcus pyogenes dCas9 (3xFLAG-dCas9) and retrovirally transduced gRNA into primary CD4+ T cells from these mice for enChIP. Using this approach, we achieved high yields of enChIP at the targeted genomic region. Our novel transgenic mouse lines provide a valuable tool for enChIP analysis in primary mouse cells.
DC.rights	© 2018 The Authors. Genes to Cells published by Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd. This article is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
DC.format	78574
citation_title	Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis
citation_author	Fujita, Toshitsugu
	Kitaura, Fusako
	Oji, Asami
	Tanigawa, Naoki
	Yuno, Miyuki
	Ikawa, Masahito
	Taniuchi, Ichiro
	Fujii, Hodaka
citation_publisher	Blackwell Publishing Ltd
citation_language	English
citation_date	2018-04-10
citation_journal_title	Genes to Cells
citation_volume	23
citation_issue	4
citation_firstpage	318
citation_lastpage	325
citation_doi	https://doi.org/10.1111/gtc.12573
citation_pmid	29480524
citation_public_url	https://hdl.handle.net/11094/78574

Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis

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