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. Author manuscript; available in PMC: 2016 Jul 1.
Published in final edited form as: Nat Biotechnol. 2015 Dec 7;34(1):78–83. doi: 10.1038/nbt.3439

Figure 1. Gene disruption by homology directed repair at three separate loci causes recessive female sterility.

Figure 1

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.