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. 2018 Jan 18;8(3):999–1018. doi: 10.1534/g3.117.300557

Figure 7.

Figure 7

Modulation of gene drive activity by titration of Cas9 and nuclear shuttling. (A) Gene drives were generated based on the 16 plasmid-borne Cas9 constructs from Figure 3 and integrated at the HIS3 locus. All gene drive strains (GFY-2751–GFY-2766) were transformed with the sgRNA(u1) 20 bp WT guide plasmid and mated to the two target strains (GFY-3206 and GFY-3207). Following diploid selection and preinduction in a raffinose/sucrose mixture, diploid yeast were cultured in YP + galactose for 1.25, 2.5, or 5.0 hr prior to plating. Representative plates (the Cas9-eGFP fusion number illustrated for clarity) for two groupings are illustrated at the 5 hr time point on SD-LEU and SD-HIS medium (left). The percentage of yeast with active gene drives (percentage of colonies dead on SD-HIS) was quantified in triplicate (right). Error, SD. (B) Two-way comparisons between strains from (A) were performed using an unpaired t-test. Red text, p-values > 0.05. Asterisk, the collective average of all three strains was used for comparisons. (C) The data from (A) was reordered from least to greatest percentage of active gene drive (top). The data from (A) is presented in a histogram with 10% binning categories (bottom).