Diphtheria toxin-mediated transposon-driven poly (A)-trapping efficiently disrupts transcriptionally silent genes in embryonic stem cells

Bai, Jie; Kondo, Ryohei; Mayasari, N. Ika; Shigeoka, Toshiaki; Isotani, Ayako; Ikawa, Masahito; Sashida, Goro; Kawaichi, Masashi; Ishida, Yasumasa

doi:https://doi.org/10.1002/dvg.23386

Diphtheria toxin-mediated transposon-driven poly (A)-trapping efficiently disrupts transcriptionally silent genes in embryonic stem cells

Bai, Jie; Kondo, Ryohei; Mayasari, N. Ika et al.

Genesis, 2020, 58(9), e23386

アクセス数:133件（2025-04-18 02:32 集計）

固定URL: https://hdl.handle.net/11094/78542

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タイトル	Diphtheria toxin-mediated transposon-driven poly (A)-trapping efficiently disrupts transcriptionally silent genes in embryonic stem cells
著者	NRID 1000060775336 1000060775336 著者名 Bai, Jie
	著者名 Kondo, Ryohei
	著者名 Mayasari, N. Ika
	NRID 1000040457148 1000040457148 著者名 Shigeoka, Toshiaki
	NRID 1000020444523 1000020444523 著者名 Isotani, Ayako
	NRID 1000020304066 1000020304066 ORCID 0000-0001-9859-6217 0000-0001-9859-6217 researchmap permalink read0054910 read0054910 Scopus Author ID 16689054500 16689054500 著者名 Ikawa, Masahito
	NRID 1000070349447 1000070349447 著者名 Sashida, Goro
	NRID 1000000195041 1000000195041 著者名 Kawaichi, Masashi
	NRID 1000010221756 1000010221756 著者名 Ishida, Yasumasa
キーワード等	cell lineage ablation
	diphtheria toxin
	embryonic stem cells
	gene trap
	Tol2 transposon
内容	Bai, J, Kondo, R, Mayasari, NI, et al. Diphtheria toxin‐mediated transposon‐driven poly (A)‐trapping efficiently disrupts transcriptionally silent genes in embryonic stem cells. genesis. 2020; 58:e23386. https://doi.org/10.1002/dvg.23386
抄録	Random gene trapping is the application of insertional mutagenesis techniques that are conventionally used to inactivate protein-coding genes in mouse embryonic stem (ES) cells. Transcriptionally silent genes are not effectively targeted by conventional random gene trapping techniques, thus we herein developed an unbiased poly (A) trap (UPATrap) method using a Tol2 transposon, which preferentially integrated into active genes rather than silent genes in ES cells. To achieve efficient trapping at transcriptionally silent genes using random insertional mutagenesis in ES cells, we generated a new diphtheria toxin (DT)-mediated trapping vector, DTrap that removed cells, through the expression of DT that was induced by the promoter activity of the trapped genes, and selected trapped clones using the neomycin-resistance gene of the vector. We found that a double-DT, the dDT vector, dominantly induced the disruption of silent genes, but not active genes, and showed more stable integration in ES cells than the UPATrap vector. The dDT vector disrupted differentiated cell lineage genes, which were silent in ES cells, and labeled trapped clone cells by the expression of EGFP upon differentiation. Thus, the dDT vector provides a systematic approach to disrupt silent genes and examine the cellular functions of trapped genes in the differentiation of target cells and development.
出版者	John Wiley and Sons Inc
収録物名	Genesis
巻(号)	58(9)
ページ	e23386
刊行年月	2020-09-16
言語	英語
ハンドルURL	https://hdl.handle.net/11094/78542
PISSN	1526954X
eISSN	1526968X
NCID	AA11620250
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.1002/dvg.23386
関連情報 (isIdenticalTo)	PMID 32645254
アクセス権	open access
権利情報	© 2020 The Authors. Genesis published by Wiley Periodicals LLC. This article is licensed under a Creative Commons Attribution 4.0 International License.
出版タイプ	Version of Record
カテゴリ	学術雑誌論文 Journal Article

資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
DCTERMS.bibliographicCitation	Genesis. 2020, 58(9), p. e23386
DC.title	Diphtheria toxin-mediated transposon-driven poly (A)-trapping efficiently disrupts transcriptionally silent genes in embryonic stem cells
DC.creator	Bai, Jie
	Kondo, Ryohei
	Mayasari, N. Ika
	Shigeoka, Toshiaki
	Isotani, Ayako
	Ikawa, Masahito
	Sashida, Goro
	Kawaichi, Masashi
	Ishida, Yasumasa
DC.publisher	John Wiley and Sons Inc
DC.language' scheme='DCTERMS.RFC1766	英語
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2020-09-16
DC.identifier	https://doi.org/10.1002/dvg.23386
DC.subject	cell lineage ablation
	diphtheria toxin
	embryonic stem cells
	gene trap
	Tol2 transposon
DC.description	Bai, J, Kondo, R, Mayasari, NI, et al. Diphtheria toxin‐mediated transposon‐driven poly (A)‐trapping efficiently disrupts transcriptionally silent genes in embryonic stem cells. genesis. 2020; 58:e23386. https://doi.org/10.1002/dvg.23386
DCTERMS.abstract	Random gene trapping is the application of insertional mutagenesis techniques that are conventionally used to inactivate protein-coding genes in mouse embryonic stem (ES) cells. Transcriptionally silent genes are not effectively targeted by conventional random gene trapping techniques, thus we herein developed an unbiased poly (A) trap (UPATrap) method using a Tol2 transposon, which preferentially integrated into active genes rather than silent genes in ES cells. To achieve efficient trapping at transcriptionally silent genes using random insertional mutagenesis in ES cells, we generated a new diphtheria toxin (DT)-mediated trapping vector, DTrap that removed cells, through the expression of DT that was induced by the promoter activity of the trapped genes, and selected trapped clones using the neomycin-resistance gene of the vector. We found that a double-DT, the dDT vector, dominantly induced the disruption of silent genes, but not active genes, and showed more stable integration in ES cells than the UPATrap vector. The dDT vector disrupted differentiated cell lineage genes, which were silent in ES cells, and labeled trapped clone cells by the expression of EGFP upon differentiation. Thus, the dDT vector provides a systematic approach to disrupt silent genes and examine the cellular functions of trapped genes in the differentiation of target cells and development.
DC.rights	© 2020 The Authors. Genesis published by Wiley Periodicals LLC. This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	78542
citation_title	Diphtheria toxin-mediated transposon-driven poly (A)-trapping efficiently disrupts transcriptionally silent genes in embryonic stem cells
citation_author	Bai, Jie
	Kondo, Ryohei
	Mayasari, N. Ika
	Shigeoka, Toshiaki
	Isotani, Ayako
	Ikawa, Masahito
	Sashida, Goro
	Kawaichi, Masashi
	Ishida, Yasumasa
citation_publisher	John Wiley and Sons Inc
citation_language	英語
citation_date	2020-09-16
citation_journal_title	Genesis
citation_volume	58
citation_issue	9
citation_firstpage	e23386
citation_doi	https://doi.org/10.1002/dvg.23386
citation_pmid	32645254
citation_public_url	https://hdl.handle.net/11094/78542

Diphtheria toxin-mediated transposon-driven poly (A)-trapping efficiently disrupts transcriptionally silent genes in embryonic stem cells

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