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

Author:

Tanaka Masayuki,Yokoyama Keiko,Hayashi Hideki,Isaki Sanae,Kitatani Kanae,Wang Ting,Kawata Hisako,Matsuzawa Hideyuki,Gurumurthy Channabasavaiah B.,Miura Hiromi,Ohtsuka MasatoORCID

Abstract

AbstractCRISPR 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-Knock-ins and Random Inserts Searching PRotocol), 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.

Funder

Research and Study Project of Tokai University General Research Organization

Tokai University School of Medicine Project Research

National Human Genome Research Institute

National Institute of General Medical Sciences

Publisher

Springer Science and Business Media LLC

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