Fig. 4

Mutant alleles identified from the G₁ population at the shi^ts1 locus. Shi^ts1 CRISPR/Cas9 mutagenesis experiments produced two mutant genotypes (the shi^ts1 single base substitution and a 4 bp deletion) in G₁ progeny. The shi^ts1 G to A mutant allele (highlighted in red) causes a glycine (G) to aspartic acid (D) amino acid change (highlighted in blue). A 4 bp deletion (indicated by the red dotted lines) causes a frameshift and consequently a change in the downstream amino acid sequence (highlighted in blue). WT = wild type (reference genome) sequence, KI = knock-in, Del = deletion. The 3 bp PAM sequence is underlined with a bold line and the 20 bp guide sequence with dotted lines, with the red arrow indicating the PAM cut site. The base where the shi^ts1 point mutation is made is highlighted in bold and with an asterisk