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-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.
Mice, Animals, Clustered Regularly Interspaced Short Palindromic Repeats, Gene Knock-In Techniques, CRISPR-Cas Systems, DNA, Genome, Gene Editing
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Tanaka, Masayuki; Yokoyama, Keiko; Hayashi, Hideki; Isaki, Sanae; Kitatani, Kanae; Wang, Ting; Kawata, Hisako; Matsuzawa, Hideyuki; Gurumurthy, Channabasavaiah B.; Miura, Hiromi; and Ohtsuka, Masato, "CRISPR-KRISPR: A Method to Identify On-Target and Random Insertion of Donor DNAs and their Characterization in Knock-In Mice" (2022). Journal Articles: Pharmacology & Experimental Neuroscience. 62.