Figure 4. Modified donors promote precise editing in Caenorhabditis elegans.

(A) Schematic showing end-modified double-stranded DNA (dsDNA) donors (25 ng/µl) with short (~35 bp) homology arms to insert gfp tag. (B) Number of green fluorescent protein (GFP) expressing animals among entire F1 brood of two representative P0 animals for each donor type are plotted for eft-3p reporter locus. Fraction of F1 animals expressing GFP among (C) Roller and (D) non-Roller cohorts are plotted as percentage for eft-3p locus. Similarly, (E) number of GFP expressing animals among two representative broods, fraction of F1 animals expressing GFP among (F) Roller and (G) non-Roller cohorts are plotted for csr-1 locus. Open bars (Rollers) and closed bars represent (non-Rollers) median. Number of GFP expressing animals among total number of animals scored per cohort are shown above the bars. n ≥ 4 broods for each donor condition. p-Values were calculated using one-way ANOVA and in all cases end-modified donors were compared to unmodified donors (Tukey’s multiple comparisons test; ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05; ns, not significant).

Figure 4—figure supplement 1. Precise insertions are germline transmitted in Caenorhabditis elegans.

(A) Schematic representation of the eft3p-gfp (partial) locus edited with double-stranded DNA (dsDNA) donors with or without end modifications in C. elegans. Precisely edited animals express green fluorescent protein (GFP) signal ubiquitously. (B) GFP-positive F1 animals were cloned, and their progeny (F2s) were scored for GFP expression. Number of F1s that produced GFP expressing F2 in a Mendelian fashion are shown under F2 transmission column.