Figure 1. Gene disruption by homology directed repair at three separate loci causes recessive female sterility.
A) A plasmid-based source of either a TALEN or Cas9 coupled with a gRNA induces a double stranded break at the _target locus. A plasmid (hdrGFP) containing regions of homology immediately upstream and downstream of the cut site acts as a template for homology-directed repair. Internal to the homology regions a 3xP3::GFP cassette identifies hdrGFP integration events and two attP sites facilitate secondary modification of the locus through RMCE. B) PCR was used to confirm the _targeted loci in wild type individuals as well as those homozygous and heterozygous for the hdrGFP allele. The primer pair used is indicated in A (blue arrows). C) Counts of larval progeny from individual females homozygous or heterozygous for hdrGFP alleles mated to wild type males. Heterozygous docking lines for all 3 loci showed at least full fertility compared to wild type females. A minimum of 20 individuals were tested for each line. Vertical bars represent the mean and error bars the standard error of the mean.