Fig. 5: Impaired homology arms affect drive efficiency.

a Inheritance rates of the three modified w-[y1,w2] gRNA lines (combined with vasa-Cas9 in yellow) in which the homology (b) was impaired by removing 20 bp of sequence on both sides of the transgene (both sides), only on the PAM-proximal side (PAM proximal), or on the PAM-distal side (PAM distal). b Genomic and transgene sequences, 20 base pairs impaired homology sequences indicated in black, blue, and magenta; blue: gRNA target, magenta: PAM sequence, and green triangles highlight the cut location and gRNA-GFP transgene insertion site. GFP transgene is shown in green. Dashes highlight the missing sequences in the impaired homology constructs. The green (gRNA) and red (Cas9) markings on the y-axis in (a) indicate the average inheritance value of the control cross, with perfect homology, from Fig. 2a, e. c–f A schematic of each construct showing how the homology is impaired during the repair process. The bottom portion of the panel contains a 2D-correlation plot in which the inheritance of the DsRed-marked construct (Cas9) is represented on the x-axis, while the inheritance of the GFP-gRNA cassette is indicated on the y-axis. The individual data points are overlaid on a 2D-density distribution plot, and a 1D distribution of the data on each axis is also shown on the top and to he right of each graph. (c) Control: 2D-correlation graph of the data from Fig. 2 (perfect homology in both transgenes) representing the baseline. d Homology impaired on both sides of the gRNA transgene reduces allelic conversion of the GFP-marked construct. e Impaired homology on the PAM-proximal side behaves similarly to the control. f Impaired homology on the PAM-distal side seems to be slightly affected in terms of GFP inheritance, as individual data points are distributed almost equally on both sides of the diagonal. c–e Quantification of the percentage of data points falling over (green), on (black), and under (red) the midline is overlaid on each graph. Raw phenotypical scoring is provided as “Supplementary Data 4”.