Double strand break repair by capture of retrotransposon sequences and reverse-transcribed spliced mRNA sequences in mouse zygotes

Ono, Ryuichi; Ishii, Masayuki; Fujihara, Yoshitaka; Kitazawa, Moe; Usami, Takako; Kaneko, Tomoko; Kanno, Jun; Ikawa, Masahito; Ishino, Fumitoshi

doi:https://doi.org/10.1038/srep12281

Double strand break repair by capture of retrotransposon sequences and reverse-transcribed spliced mRNA sequences in mouse zygotes

Ono, Ryuichi; Ishii, Masayuki; Fujihara, Yoshitaka et al.

Scientific Reports, 2015, 5, 12281

アクセス数:216件（2025-04-18 03:07 集計）

固定URL: https://hdl.handle.net/11094/78597

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タイトル	Double strand break repair by capture of retrotransposon sequences and reverse-transcribed spliced mRNA sequences in mouse zygotes
著者	NRID 1000010401358 1000010401358 著者名 Ono, Ryuichi
	著者名 Ishii, Masayuki
	NRID 1000070578848 1000070578848 著者名 Fujihara, Yoshitaka
	NRID 1000040801965 1000040801965 著者名 Kitazawa, Moe
	著者名 Usami, Takako
	NRID 1000020221757 1000020221757 著者名 Kaneko, Tomoko 著者の別表記 Ishino, Tomoko
	NRID 1000090186172 1000090186172 著者名 Kanno, Jun
	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 1000060159754 1000060159754 著者名 Ishino, Fumitoshi
内容	Ono, R., Ishii, M., Fujihara, Y. et al. Double strand break repair by capture of retrotransposon sequences and reverse-transcribed spliced mRNA sequences in mouse zygotes. Sci Rep 5, 12281 (2015). https://doi.org/10.1038/srep12281
抄録	The CRISPR/Cas system efficiently introduces double strand breaks (DSBs) at a genomic locus specified by a single guide RNA (sgRNA). The DSBs are subsequently repaired through non-homologous end joining (NHEJ) or homologous recombination (HR). Here, we demonstrate that DSBs introduced into mouse zygotes by the CRISPR/Cas system are repaired by the capture of DNA sequences deriving from retrotransposons, genomic DNA, mRNA and sgRNA. Among 93 mice analysed, 57 carried mutant alleles and 22 of them had long de novo insertion(s) at DSB-introduced sites; two were spliced mRNAs of Pcnt and Inadl without introns, indicating the involvement of reverse transcription (RT). Fifteen alleles included retrotransposons, mRNAs, and other sequences without evidence of RT. Two others were sgRNAs with one containing T7 promoter-derived sequence suggestive of a PCR product as its origin. In conclusion, RT-product-mediated DSB repair (RMDR) and non-RMDR repair were identified in the mouse zygote. We also confirmed that both RMDR and non-RMDR take place in CRISPR/Cas transfected NIH-3T3 cells. Finally, as two de novo MuERV-L insertions in C57BL/6 mice were shown to have characteristic features of RMDR in natural conditions, we hypothesize that RMDR contributes to the emergence of novel DNA sequences in the course of evolution.
出版者	Nature Research
収録物名	Scientific Reports
巻(号)	5
ページ	12281
刊行年月	2015-07-28
言語	英語
ハンドルURL	https://hdl.handle.net/11094/78597
eISSN	20452322
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.1038/srep12281
関連情報 (isIdenticalTo)	PMID 26216318
アクセス権	open access
権利情報	© 2015 The Author(s). 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	Scientific Reports. 2015, 5, p. 12281
DC.title	Double strand break repair by capture of retrotransposon sequences and reverse-transcribed spliced mRNA sequences in mouse zygotes
DC.creator	Ono, Ryuichi
	Ishii, Masayuki
	Fujihara, Yoshitaka
	Kitazawa, Moe
	Usami, Takako
	Kaneko, Tomoko
	Kanno, Jun
	Ikawa, Masahito
	Ishino, Fumitoshi
DC.creator	Ishino, Tomoko
DC.publisher	Nature Research
DC.language' scheme='DCTERMS.RFC1766	英語
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2015-07-28
DC.identifier	https://doi.org/10.1038/srep12281
DC.description	Ono, R., Ishii, M., Fujihara, Y. et al. Double strand break repair by capture of retrotransposon sequences and reverse-transcribed spliced mRNA sequences in mouse zygotes. Sci Rep 5, 12281 (2015). https://doi.org/10.1038/srep12281
DCTERMS.abstract	The CRISPR/Cas system efficiently introduces double strand breaks (DSBs) at a genomic locus specified by a single guide RNA (sgRNA). The DSBs are subsequently repaired through non-homologous end joining (NHEJ) or homologous recombination (HR). Here, we demonstrate that DSBs introduced into mouse zygotes by the CRISPR/Cas system are repaired by the capture of DNA sequences deriving from retrotransposons, genomic DNA, mRNA and sgRNA. Among 93 mice analysed, 57 carried mutant alleles and 22 of them had long de novo insertion(s) at DSB-introduced sites; two were spliced mRNAs of Pcnt and Inadl without introns, indicating the involvement of reverse transcription (RT). Fifteen alleles included retrotransposons, mRNAs, and other sequences without evidence of RT. Two others were sgRNAs with one containing T7 promoter-derived sequence suggestive of a PCR product as its origin. In conclusion, RT-product-mediated DSB repair (RMDR) and non-RMDR repair were identified in the mouse zygote. We also confirmed that both RMDR and non-RMDR take place in CRISPR/Cas transfected NIH-3T3 cells. Finally, as two de novo MuERV-L insertions in C57BL/6 mice were shown to have characteristic features of RMDR in natural conditions, we hypothesize that RMDR contributes to the emergence of novel DNA sequences in the course of evolution.
DC.rights	© 2015 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	78597
citation_title	Double strand break repair by capture of retrotransposon sequences and reverse-transcribed spliced mRNA sequences in mouse zygotes
citation_author	Ono, Ryuichi
	Ishii, Masayuki
	Fujihara, Yoshitaka
	Kitazawa, Moe
	Usami, Takako
	Kaneko, Tomoko
	Kanno, Jun
	Ikawa, Masahito
	Ishino, Fumitoshi
citation_publisher	Nature Research
citation_language	英語
citation_date	2015-07-28
citation_journal_title	Scientific Reports
citation_volume	5
citation_firstpage	12281
citation_doi	https://doi.org/10.1038/srep12281
citation_pmid	26216318
citation_public_url	https://hdl.handle.net/11094/78597

Double strand break repair by capture of retrotransposon sequences and reverse-transcribed spliced mRNA sequences in mouse zygotes

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