Figure 4: Increased transfection efficiency uncovers homology-mediated RI events.

(a) Comparison of transfection efficiency between the three transfection methods. Transient expression assays were performed using a luciferase (left) or GFP (right) expression vector (see Supplementary Fig. 2l,m). (b) Integration frequency of MaxCyte transfected pPGKneo in wild-type, LIG4^−/−, LIG4^−/−POLQ^−/− and LIG4-complemented LIG4^−/−POLQ^−/− cells. (c) Junction sequence in a rare RI clone obtained from LIG4^−/−POLQ^−/− cells. (d) Gene-targeting efficiency, targeted integration frequency, and RI frequency of MaxCyte transfected pHPRT8.9-Neo2(−) in wild-type, LIG4^−/−, LIG4^−/−POLQ^−/− and LIG4-complemented LIG4^−/−POLQ^−/− (#3) cells. (e) Junction sequences in rare RI clones obtained from LIG4^−/−POLQ^−/− cells.