Figure 1. Dual sgRNA-guided deletion of the rde-12 gene.

(A) Schematic of the screen for CRISPR/Cas9 genome editing events. The dominant transformation marker mCherry was co-injected with Cas9 and sgRNA #1 and #2 expression plasmids. F1 animals with mCherry expression were grown on unc-15 RNAi, and phenotypes of F2 were scored. The RNAi suppressors were PCR amplified and sequenced. (B) Schematic of the rde-12 gene. Positions of sgRNA-guided cleavage sites are indicated. (C) Summary of microinjection experiments. (D) Sequence alignments of the rde-12 gene in wild-type and mutant animals. The PAM sequence is labeled in red and overlined. Dash indicates deletion. Lowercase indicates insertion. The numbers in parentheses within the sequence represent the number of bases not shown. The number of deleted (−) or inserted (+) bases is shown on the right of each indel. Numbers on the top of sequences indicate positions relative to the transcription start site.