Additional file 3 of Monitoring autochthonous lung tumors induced by somatic CRISPR gene editing in mice using a secreted luciferase

Additional file 3: sgRNA off-target analysis. Trp53, Rb1 and Rbl2 sgRNAs were analyzed for potential off-targets using the CRISPRoff Tool version 1.2. Shown are the Top10 hits with an intragenic location and “NGG” PAM. The Top10 hits for each of the Trp53, Rb1 and Rbl2 sgRNAs were PCR amplified using genomic tumor DNA from 3 AV-PR.Cre and 3 AV-PRL.Cre infected mice. PCR amplicons were analyzed for mutations by Sanger sequencing. Results are shown color-coded. Identified on-target and off-target mutations are shown in detail. Off-target mutations were all heterozygous missense mutations.