NDLI: Dual sgRNAs facilitate CRISPR/Cas9-mediated mouse genome targeting.

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Dual sgRNAs facilitate CRISPR/Cas9-mediated mouse genome targeting.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Zhou, Jiankui Wang, Jianying Shen, Bin Chen, Li Su, Yang Yang, Jing Zhang, Wensheng Tian, Xuemei Huang, Xingxu
Description	Country affiliation: China Author Affiliation: Zhou J ( MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, National Resource Center for Mutant Mice, Nanjing, China.)
Abstract	The bacterial clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system is a versatile RNA-guided mammalian genome modification system. One-step generation of mouse genome targeting has been achieved by co-microinjection of one-cell stage embryos with Cas9 mRNA and small/single guide (sg)RNA. Many studies have focused on enhancing the efficiency of this system. In the present study, we report that simultaneous use of dual sgRNAs to target an individual gene significantly improved the Cas9-mediated genome targeting with a bi-allelic modification efficiency of up to 78%. We further observed that the target gene modifications were characterized by efficient germline transmission and site-dependent off-target effects, and also that the apolipoprotein E gene knockout-mediated defects in blood biochemical parameters were recapitulated by CRISPR/Cas9-mediated heritable gene modification. Our results provide a dual sgRNAs strategy to facilitate CRISPR/Cas9-mediated mouse genome targeting.
File Format	HTM / HTML
ISSN	1742464X
Issue Number	7
Journal	FEBS Journal
Volume Number	281
e-ISSN	17424658
Language	English
Publisher	Wiley (on behalf of the Federation of European Biochemical Societies)
Publisher Date	2014-04-01
Publisher Place	Great Britain (UK)
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Biochemistry Crispr-cas Systems Gene Targeting Methods Micrornas Metabolism Animals Mice Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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Similar Documents

Editing the Mouse Genome Using the CRISPR-Cas9 System.

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