Creation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the CRISPR/Cas9 system

Abstract The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a useful tool for creation of mutant mice with mutations mirroring those in human patients. Various methods have been developed for this purpose, including deletions, inversion...

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Autores principales:	Tomoko Kato, Satoshi Hara, Yuji Goto, Yuya Ogawa, Haruka Okayasu, Souichirou Kubota, Moe Tamano, Miho Terao, Shuji Takada
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/020016776a324b9792c8769dda31a05a
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spelling	oai:doaj.org-article:020016776a324b9792c8769dda31a05a2021-12-02T11:53:03ZCreation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the CRISPR/Cas9 system10.1038/s41598-017-00140-92045-2322https://doaj.org/article/020016776a324b9792c8769dda31a05a2017-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00140-9https://doaj.org/toc/2045-2322Abstract The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a useful tool for creation of mutant mice with mutations mirroring those in human patients. Various methods have been developed for this purpose, including deletions, inversions, and translocations. So far, mutant mice with deletions of up to 1.2 megabases (Mb) have been generated by microinjection of the CRISPR/Cas9 system into fertilized eggs; however, a method for generation of mutant mice with a deletion of more than several Mb size is necessary because such deletions have often been identified as possible causes of human diseases. With an aim to enable the generation of disease models carrying large deletions with a breakpoint in custom-designed sequences, we developed a method for induction of an Mb-sized deletion by microinjection of a pair of sgRNAs, Cas9, and a donor plasmid into fertilized eggs. Using this method, we efficiently and rapidly generated mutant mice carrying deletions up to 5 Mb.Tomoko KatoSatoshi HaraYuji GotoYuya OgawaHaruka OkayasuSouichirou KubotaMoe TamanoMiho TeraoShuji TakadaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tomoko Kato Satoshi Hara Yuji Goto Yuya Ogawa Haruka Okayasu Souichirou Kubota Moe Tamano Miho Terao Shuji Takada Creation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the CRISPR/Cas9 system
description	Abstract The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a useful tool for creation of mutant mice with mutations mirroring those in human patients. Various methods have been developed for this purpose, including deletions, inversions, and translocations. So far, mutant mice with deletions of up to 1.2 megabases (Mb) have been generated by microinjection of the CRISPR/Cas9 system into fertilized eggs; however, a method for generation of mutant mice with a deletion of more than several Mb size is necessary because such deletions have often been identified as possible causes of human diseases. With an aim to enable the generation of disease models carrying large deletions with a breakpoint in custom-designed sequences, we developed a method for induction of an Mb-sized deletion by microinjection of a pair of sgRNAs, Cas9, and a donor plasmid into fertilized eggs. Using this method, we efficiently and rapidly generated mutant mice carrying deletions up to 5 Mb.
format	article
author	Tomoko Kato Satoshi Hara Yuji Goto Yuya Ogawa Haruka Okayasu Souichirou Kubota Moe Tamano Miho Terao Shuji Takada
author_facet	Tomoko Kato Satoshi Hara Yuji Goto Yuya Ogawa Haruka Okayasu Souichirou Kubota Moe Tamano Miho Terao Shuji Takada
author_sort	Tomoko Kato
title	Creation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the CRISPR/Cas9 system
title_short	Creation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the CRISPR/Cas9 system
title_full	Creation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the CRISPR/Cas9 system
title_fullStr	Creation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the CRISPR/Cas9 system
title_full_unstemmed	Creation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the CRISPR/Cas9 system
title_sort	creation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the crispr/cas9 system
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/020016776a324b9792c8769dda31a05a
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Creation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the CRISPR/Cas9 system

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