FIG 3 .

Sequencing of the eGFP gene in GFP-disrupted clones sorted from parasite populations transfected with eGFP-targeting sgRNA 3 (top) and sgRNA 1 (bottom). Underlined is the sgRNA targeting sequence, the PAM is boxed (on the reverse strand), red marks show microhomology sequences flanking the sgRNA targeting sequence, and double-stranded cut sites are indicated with blue arrows. The sequence of negative clone 1 in the sgRNA-1 set (bottom) shows a single-base deletion in this region, likely due to a spontaneous mutation in GFP. All other sequences showed a 33-bp deletion; although deletion junctions occurred at slightly different positions, they all fell between a pair of homologous sequences of 10 bp flanking the cut site, as predicted by repair via an MMEJ pathway.