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

Number of Access:774（2025-08-22 01:08 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/78597

Link to primary information

File	Format	Terms of use	Size	Views	Date.Available	Description	information
SciRep_05_12281	pdf	None	3.76 MB	111	2021.02.08

Creative Commons : Attribution

Item Information

Output File Export EndNote Basic Export Mendeley

Title	Double strand break repair by capture of retrotransposon sequences and reverse-transcribed spliced mRNA sequences in mouse zygotes
Creator	NRID 1000010401358 1000010401358 CreatorName Ono, Ryuichi
	CreatorName Ishii, Masayuki
	NRID 1000070578848 1000070578848 CreatorName Fujihara, Yoshitaka
	NRID 1000040801965 1000040801965 CreatorName Kitazawa, Moe
	CreatorName Usami, Takako
	NRID 1000020221757 1000020221757 CreatorName Kaneko, Tomoko Creator.Alternative Ishino, Tomoko
	NRID 1000090186172 1000090186172 CreatorName Kanno, Jun
	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 1000060159754 1000060159754 CreatorName Ishino, Fumitoshi
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
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.
Publisher	Nature Research
Source Title	Scientific Reports
Volume (Issue)	5
Page	12281
Date of Issued	2015-07-28
Language	English
Handle URL	https://hdl.handle.net/11094/78597
eISSN	20452322
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1038/srep12281
Relation.isIdenticalTo	PMID 26216318
Access Rights	open access
CopyRight	© 2015 The Author(s). This article is licensed under a Creative Commons Attribution 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	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	English
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	English
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

Link to primary information

Item Information

Show details