Fig. 4. Sanger sequencing results show various indel mutations in the selected UCHL-1 knockout clones.

Genomic PCR products from the selected iPSC clones were subcloned in a T-vector, and more than fifty individual colonies were analyzed for sequencing. Genotypes are listed as either homozygous, indicating that both alleles have the same mutation, or heterozygous, indicating that there is a mutation in at least one allele and the other allele contains a different mutation. Specific nucleotide sequences of insertions (underlined uppercases, green), deletions (red dash) and substitutions (lower cases, blue or black) around the editing site are indicated. The guide RNA sequence is underlined, and the PAM sequence is indicated in black boxes in the wt sequence. Additional marked sequences* show detected sequences that were not characterized for allelic features of homo or heterozygosity of the clones.