Fig. 4

Analysis of the potential off-target effects in the genome of TRP53 R172P KI mice with PCR and Sanger sequencing. a BLAST of gRNA applied in the study with mouse genome showed 23 matching loci with the Trp53 locus on the top. b The top 10 putative off-target loci in genome were PCR amplified with their own specific primer pairs and separated with electrophoresis on agarose gel. c Sanger Sequencing of the target PCR products (short band) of #1 off-target locus (Trp53 pseudogene) showed overlapping peaks in KI1 and KI3 mice but not in WT and KI2 mice. BLAST of the PCR products of #1 off-target locus (Trp53 pseudogene) showed tons of mismatches since the gRNA region in KI1 and KI3 mice but not in WT and KI2 mice, indicating the off-target effects of locus #1 in KI1 and KI3 mice. The results of KI2 and KI3 are shown on Additional file 1: Figure S1