biostudies-literatureTanaka MNHGRI NIH HHSResearch and Study Project of Tokai University General Research OrganizationNational Institute of General Medical SciencesNational Human Genome Research InstituteNIGMS NIH HHSTokai University School of Medicine Project Research228https://www.ebi.ac.uk/biostudies/studies/S-EPMC9594901biostudies-literatureUnknown23(1)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.Genome biologyCRISPR-KRISPR: a method to identify on-target and random insertion of donor DNAs and their characterization in knock-in mice.PMC95949012016-20172016-2018R21GM129559R35HG010719Miura HHayashi HIsaki SGurumurthy CBMatsuzawa HWang TKawata HKitatani KYokoyama KOhtsuka MTanaka MfalseCRISPR-KRISPR: a method to identify on-target and random insertion of donor DNAs and their characterization in knock-in mice.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.2022-01-01T00:00:00Z2022 Oct2025-04-18T11:42:26.14Z2024-10-15T01:22:02.543ZS-EPMC95949013628431110.1186/s13059-022-02779-8