Fig. 2: Generation of a Y chromosome-linked genome editor and evaluation of Y-linked Cas9 activity.

a Genetic constructs used to generate a YLE based on Cas9 expression under the germline vas2 promoter, targeting the female isoform of the doublesex gene. The constructs contained a gene coding for either CFP or RFP under the 3xP3 promoter and were inserted in the Y-linked loci on the YAttP and the Ygfp strains using integrase-mediated recombination. b Schematic representation of the An. gambiae doublesex gene, showing the male isoform (top) and the female isoform (bottom). Boxes represent exons, and lines represent introns. CDS and UTRs are shown in dark and light colours, respectively. Introns are not drawn to scale. The red arrow indicates the site targeted by the YLE. c Illustration of homology-directed repair (homing) with a modified dsx allele (dsxF^GFP-null) used to evaluate the Cas9 activity of the YLE^dsx strains. d Percentage of progeny from YLE^dsx-A males (dark green) and YLE^dsx-B males (light green) that inherited the dsxF^GFP-null allele when crossed to wild-type individuals (n ≥ 30). Each dot corresponds to the progeny of a different mating couple. The dashed line indicates the expected Mendelian inheritance. Numbers on the right indicate the mean, the s.e.m., and the size of each sample (n). Source data are provided as a Source Data file.