Precise genome-wide base editing by the CRISPR Nickase system in yeast

Abstract The CRISPR/Cas9 system has been applied to efficient genome editing in many eukaryotic cells. However, the bases that can be edited by this system have been limited to those within the protospacer adjacent motif (PAM) and guide RNA-targeting sequences. In this study, we developed a genome-w...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Atsushi Satomura, Ryosuke Nishioka, Hitoshi Mori, Kosuke Sato, Kouichi Kuroda, Mitsuyoshi Ueda
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/257428f6c7f3431591977f55616b99b8
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:257428f6c7f3431591977f55616b99b8
record_format	dspace
spelling	oai:doaj.org-article:257428f6c7f3431591977f55616b99b82021-12-02T16:08:21ZPrecise genome-wide base editing by the CRISPR Nickase system in yeast10.1038/s41598-017-02013-72045-2322https://doaj.org/article/257428f6c7f3431591977f55616b99b82017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02013-7https://doaj.org/toc/2045-2322Abstract The CRISPR/Cas9 system has been applied to efficient genome editing in many eukaryotic cells. However, the bases that can be edited by this system have been limited to those within the protospacer adjacent motif (PAM) and guide RNA-targeting sequences. In this study, we developed a genome-wide base editing technology, “CRISPR Nickase system” that utilizes a single Cas9 nickase. This system was free from the limitation of editable bases that was observed in the CRISPR/Cas9 system, and was able to precisely edit bases up to 53 bp from the nicking site. In addition, this system showed no off-target editing, in contrast to the CRISPR/Cas9 system. Coupling the CRISPR Nickase system with yeast gap repair cloning enabled the construction of yeast mutants within only five days. The CRISPR Nickase system provides a versatile and powerful technology for rapid, site-specific, and precise base editing in yeast.Atsushi SatomuraRyosuke NishiokaHitoshi MoriKosuke SatoKouichi KurodaMitsuyoshi UedaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Atsushi Satomura Ryosuke Nishioka Hitoshi Mori Kosuke Sato Kouichi Kuroda Mitsuyoshi Ueda Precise genome-wide base editing by the CRISPR Nickase system in yeast
description	Abstract The CRISPR/Cas9 system has been applied to efficient genome editing in many eukaryotic cells. However, the bases that can be edited by this system have been limited to those within the protospacer adjacent motif (PAM) and guide RNA-targeting sequences. In this study, we developed a genome-wide base editing technology, “CRISPR Nickase system” that utilizes a single Cas9 nickase. This system was free from the limitation of editable bases that was observed in the CRISPR/Cas9 system, and was able to precisely edit bases up to 53 bp from the nicking site. In addition, this system showed no off-target editing, in contrast to the CRISPR/Cas9 system. Coupling the CRISPR Nickase system with yeast gap repair cloning enabled the construction of yeast mutants within only five days. The CRISPR Nickase system provides a versatile and powerful technology for rapid, site-specific, and precise base editing in yeast.
format	article
author	Atsushi Satomura Ryosuke Nishioka Hitoshi Mori Kosuke Sato Kouichi Kuroda Mitsuyoshi Ueda
author_facet	Atsushi Satomura Ryosuke Nishioka Hitoshi Mori Kosuke Sato Kouichi Kuroda Mitsuyoshi Ueda
author_sort	Atsushi Satomura
title	Precise genome-wide base editing by the CRISPR Nickase system in yeast
title_short	Precise genome-wide base editing by the CRISPR Nickase system in yeast
title_full	Precise genome-wide base editing by the CRISPR Nickase system in yeast
title_fullStr	Precise genome-wide base editing by the CRISPR Nickase system in yeast
title_full_unstemmed	Precise genome-wide base editing by the CRISPR Nickase system in yeast
title_sort	precise genome-wide base editing by the crispr nickase system in yeast
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/257428f6c7f3431591977f55616b99b8
work_keys_str_mv	AT atsushisatomura precisegenomewidebaseeditingbythecrisprnickasesysteminyeast AT ryosukenishioka precisegenomewidebaseeditingbythecrisprnickasesysteminyeast AT hitoshimori precisegenomewidebaseeditingbythecrisprnickasesysteminyeast AT kosukesato precisegenomewidebaseeditingbythecrisprnickasesysteminyeast AT kouichikuroda precisegenomewidebaseeditingbythecrisprnickasesysteminyeast AT mitsuyoshiueda precisegenomewidebaseeditingbythecrisprnickasesysteminyeast
_version_	1718384517500108800

Precise genome-wide base editing by the CRISPR Nickase system in yeast

Ejemplares similares