Gene editing in mouse zygotes using the CRISPR/Cas9 system

被引：37

作者：

Wefers, Benedikt ^{[1
,2
]}

Bashir, Sanum ^{[3
,4
]}

Rossius, Jana ^{[3
]}

Wurst, Wolfgang ^{[1
,2
,5
,6
]}

Kuehn, Ralf ^{[3
,4
]}

机构：

[1] German Ctr Neurodegenerat Dis DZNE, Feodor Lynen Str 17, D-81377 Munich, Germany

[2] Helmholtz Zentrum Munchen, Inst Dev Genet, German Res Ctr Environm Hlth, Ingolstadter Landstr 1, D-85764 Neuherberg, Germany

[3] Max Delbruck Ctr Mol Med, Robert Rossle Str 10, D-13125 Berlin, Germany

[4] Berlin Inst Hlth, Kapelle Ufer 2, D-10117 Berlin, Germany

[5] Tech Univ Munchen Weihenstephan, Chair Dev Genet, Helmholtz Zentrum Munchen, Ingolstadter Landstr 1, D-85764 Neuherberg, Germany

[6] Munich Cluster Syst Neurol SyNergy, Feodor Lynen Str 17, D-81377 Munich, Germany

来源：

METHODS | 2017年 / 121卷

关键词：

CRISPR; Cas9; Mouse; Zygotes; Gene editing; OFF-TARGET SITES; HUMAN-CELLS; CRISPR-CAS9; NUCLEASES; CAS9; MUTANT MICE; GUIDE RNAS; GENOME; GENERATION; SPECIFICITY; TOOL;

D O I：

10.1016/j.ymeth.2017.02.008

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The generation of targeted mouse mutants is a key technology for biomedical research. Using the CRISPR/Cas9 system for induction of targeted double-strand breaks, gene editing can be performed in a single step directly in mouse zygotes. This article covers the design of knockout and knockin alleles, preparation of reagents, microinjection or electroporation of zygotes and the genotyping of pups derived from gene editing projects. In addition we include a section for the control of experimental settings by targeting the Rosa26 locus and PCR based genotyping of blastocysts. (C) 2017 Elsevier Inc. All rights reserved.

引用

页码：55 / 67

页数：13

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