Fig. S1.

Optimization of expression level and codon use of dCas9-VPR and the sgRNA promoter. (A) Ubiquitous expression of 10XUAS-SAM, and three different 10XUAS:dCas9-VPR constructs using actin-Gal4 is not lethal, whereas expression of 20XUAS-IVS:dCas9-VPR is 100% lethal regardless of codon use. Dm, Drosophila melanogaster codon use; Hs, human codon use. (B) Expression of 20XUAS-IVS:dCas9-VPR constructs in the wing (using the ms1096-Gal4 line) was highly toxic to wing tissue in the female and caused male lethality. (C) Human codon-optimized 10UAS:dCas9-VPR led to higher levels of activation of the target gene wg than did Drosophila codon use, consistent with previous reports (8). (D) Expression of sgRNAs from UAS promoters (in the pCFD6 vector) did not lead to a higher expression than U6-driven sgRNAs (in the pCFD4 vector) in Drosophila S2R⁺ cells.