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

Masayuki Tanaka¹, Keiko Yokoyama², Hideki Hayashi²

¹Support Center for Medical Research and Education, Tokai University, Isehara, Kanagawa, 259-1193, Japan. matanaka@tokai-u.jp.

|October 26, 2022

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-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.

Keywords:

CIRCLE-seq CRISPR CRISPR-KRISPR Donor DNA Intronic region Knock-in Off target Random insertion Repeat element

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