Fig. 4.

Editing of two GFP paralogs in a single larva with a single sgRNA. The sequences from larva 4 (Fig. 3B) were clustered into eight groups using the single-nucleotide differences within an 83-bp intronic region (SI Appendix, Fig. S1). Shown are representative mutant alleles that fell into groups 1 and 6 (SI Appendix, Fig. S1), which can be assigned with confidence to GFP1 and GFP2, respectively (SI Appendix, SI Materials and Methods). Dashes, deleted nucleotides; blue lettering, inserted or altered nucleotides; red lettering, positions at which the GFP1 and GFP2 alleles in this larva differed within this 64-bp segment. (Note that only two of these positions are among those diagnostic for GFP1 vs. GFP2: cf. SI Appendix, Fig. S1.)