Fig. 1.
Prime editing in cultured S2R+ cells. (A) Diagram of PE2 expression plasmid pUAS-PE2. attB, phiC31 recombination site; NLS, nuclear localization sequence; PBS, primer-binding site; SV40, 3′ untranslated region; UAS, upstream activating sequence; w+, white+ rescue transgene. (B) Diagram of pCFD3-NS pegRNA expression plasmid. BbsI sites indicate cloning site for pegRNA encoding sequence. dU6:3, U6 promoter; U6 3′, U6 downstream region; v+, vermillion+ rescue transgene. (C) ebony genomic region showing target site and edit (ebonyG111X). (D) Dual sgRNA and pegRNA expression plasmid pCFD5-NS. tRNA, D. melanogaster and O.s. Gly tRNA sequence. (E) Schematic of S2R+ prime editing experiment. (F) Approximate quantification of precise editing and indels from S2R+ transfection experiments by amplicon sequencing. tfx, transfection.