CRISPR-KRISPR: a method to identify on-target and random insertion of donor DNAs and their characterization in knock-in mice

Abstract CRISPR tools can generate knockout and knock-in animal models easily, but the models can contain off-target genomic lesions or random insertions of donor DNAs. Simpler methods to identify off-target lesions and random insertions, using tail or earpiece DNA, are unavailable. We develop CRISP...

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Published inGenome Biology Vol. 23; no. 1; pp. 1 - 19
Main Authors Tanaka, Masayuki, Yokoyama, Keiko, Hayashi, Hideki, Isaki, Sanae, Kitatani, Kanae, Wang, Ting, Kawata, Hisako, Matsuzawa, Hideyuki, Gurumurthy, Channabasavaiah B., Miura, Hiromi, Ohtsuka, Masato
Format Journal Article
LanguageEnglish
Published London BioMed Central 25.10.2022
BMC
Subjects
Animal models
Animals
CIRCLE-seq
CRISPR
CRISPR-KRISPR
Genetic testing
Genomes
Genomics
Genotypes
Knock-in
Lesions
Method
Methods
Mutation
Off target
Random insertion
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Summary:Abstract CRISPR tools can generate knockout and knock-in animal models easily, but the models can contain off-target genomic lesions or random insertions of donor DNAs. Simpler methods to identify off-target lesions and random insertions, using tail or earpiece DNA, are unavailable. We develop CRISPR-KRISPR (CRISPR- K nock-ins and R andom I nserts S earching P R otocol), a method to identify both off-target lesions and random insertions. CRISPR-KRISPR uses as little as 3.4 μg of genomic DNA; thus, it can be easily incorporated as an additional step to genotype founder animals for further breeding.
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content type line 23
ISSN:1474-760X
1474-7596
1474-760X
DOI:10.1186/s13059-022-02779-8