Figure 7.

Efficiency of PG-specific CRISPR interference (CRISPRi) in Drosophila. A. spok_dI (= spok-dCas9) is used to ubiquitously express gRNAs targeting -110 and -482 bp upstream of dib as well as -174 and -423 bp upstream of phm (relative to transcription start site = TSS), respectively. In each case, this resulted in L3 arrest (left) and rescue to adulthood when the diet was supplemented with 20E (right). For comparison to PG-specific dib- and phm-RNAi and classic mutant phenotypes, see Figures 6A-B. B. RG-specific qPCR for dib- and phm-CRISPRi. Ring glands were dissected at 42 hr after the L2/L3 molt, three replicates per condition. * => p-value < 0.05. C. Sequences of dib and phm loci obtained from DNA of CRISPRi-treated PG nuclei show no alterations. For each condition, we sequenced 10 clones, all of which were wild type.