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Gene editing in mouse zygotes using the CRISPR/Cas9 system.
Methods 121-122, 55-67 (2017)
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.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Cas9 ; Crispr ; Gene Editing ; Mouse ; Zygotes; Off-target Sites; Human-cells; Crispr-cas9 Nucleases; Cas9 Nucleases; Mutant Mice; Guide Rnas; Genome; Generation; Specificity; Tool
ISSN (print) / ISBN
1046-2023
e-ISSN
1095-9130
Journal
Methods
Quellenangaben
Volume: 121-122,
Pages: 55-67
Publisher
Elsevier
Publishing Place
Amsterdam [u.a.]
Non-patent literature
Publications
Reviewing status
Peer reviewed
Institute(s)
Institute of Developmental Genetics (IDG)