Figure 3
Genotyping and sequence analysis of founder mice.
(a) PCR results of founder mice. Mice from two microinjection sessions were genotyped by PCR amplification and BamHI digestion. PCR and BamHI digestion products were electrophoresed in 1.2% argorose gel. Mice from the first injection session (wild type Cas9 mRNA) were labeled #1-1 to #1-10 and mice from the second injection session (mutant D10A Cas9 mRNA) were labeled #2-1 to #2-10. Wild type mouse genomic DNA was used as negative control (WT con). Cropped gels are shown. Full-length gels are provided for review in the supplementary file. (b) Sequence analysis of the mutated Fah alleles. The wild type sequence of exon5 is shown on top, with the encoded amino acids indicated above the sequence. The gRNA targeting sites are shown in red letters and PAM NGGs in turquoise. The mutant alleles of each mouse are labeled with A and B, following the mouse ID number. The indels for each mutated allele from different mice are shown in the middle. The deleted sequences are marked in gray and the inserted sequences are shown below the wild type sequence. The deletion or insertion length of each indel is shown on the right. The Fah mutant alleles with the knock-in sequence are shown in the lower panel. Only one sequence is shown for the stop codon (@) and BamHI insertion. Indel, insertion and deletion; in, insertion; del, deletion.
