Figure 2.

Genetic scheme for screening with co-CRISPR. Transgenic nos-Cas9 embryos were injected with a mix of either gRNA-e, gRNA-lbk1, and gRNA-lbk2 expression plasmids or with gRNA-e, gRNA-lbk2, and a knock-in repair template containing a FLAG-HA tag. Injected individuals were crossed to the ebony double balancer (P cross). F1 offspring were scored for e and e+. Individual F1 curly, ebony flies were crossed to a second chromosome balancer (F1 cross) to generate second chromosome lbk balanced lines. The F1 cross shown is with a single curly, ebony male. After the e broods are identified, following e is unnecessary. Homozygotes were then analyzed for mutations in lbk. Fly images were created on Genotype Builder Photoshop file S5 (Roote and Prokop 2013).