Fig. 1: Abasic sites within synthetic pegRNAs precisely block reverse transcription by M-MLV RT.
From: Modified pegRNAs mitigate scaffold-derived prime editing by-products
a Schematic representation of pegRNA scaffold capture by NHEJ in the PRINS editing assay. PRINS editing results in unintended pegRNA scaffold integrations due to flap extension past the intended reverse transcription template (RTT). b PRINS editing using modified (M) pegRNAs to prevent RT from pegRNA scaffold readthrough. c Chemical structures of modifications within rSp-pegRNA and C3-pegRNA. C96 = cytosine at the position 96 of the SpCas9 pegRNA scaffold. d Scaffold incorporation analysis by amplicon-seq of PCSK9 editing in K562 cells electroporated with PEn mRNA and unmodified pegRNA, rSp-pegRNA, or C3-pegRNA. Plots show mean ± SD of n = 3 biological replicates. e Representative CRISPResso2 alignment of PEn + pegRNA and PEn + rSp-pegRNA prime editing outcomes from experiment in Fig. 1d. Source data are provided as a Source Data file.
