Figure S4. CFTR-R785* prime editing in intestinal organoids.

(A) Guide-RNA design for the repair of the CFTR-R785* mutation in human intestinal organoids. Red bars show the nickase sites of the guide sequences and the red arrow shows the mutation site in the DNA of organoids derived from a person with cystic fibrosis. (B) pegRNA/PE3-guide pairs used in transfection for the repair of the CFTR-R785* mutation compared with adenine base editing, CRISPR/Cas9–mediated homology-dependent repair, and a negative scrambled sgRNA control. Primer-binding site length, distance to PE3 nick, and reverse transcriptase lengths are shown, as well as the mean editing efficiency. (C) Sanger sequencing traces and deconvoluted alleles of two additional prime-editing clones, one homology-dependent repair clone, and one clone repaired by adenine base editing that had been selected for by forskolin-induced swelling after transfection.