Extended Data Fig. 3: PAGE analysis and comparison of non-HPLC purified pegRNA for editing.
From: Rapid generation of long, chemically modified pegRNAs for prime editing
a. Urea-PAGE (6%) analysis of ligation products before HPLC purification. b. Urea-PAGE (6%) analysis of HPLC-purified ligated pegRNA and S-pegRNA. S-pegRNA is full-length pegRNA that was solid-phase synthesized with 3 nt chemical modifications at both ends. c–e. The efficiencies of RNP-mediated prime editing in HEK293T cells were determined by deep sequencing for 3 bp insertion at HEK3 locus (c) (n = 4), +5 G to T mutation (d) (n = 5), and 3 bp deletion (e) at VEGFA locus (PE2, n = 4, 4, 5 from left to right side; PE3, n = 4). For each electroporation, 140 pmol PE protein, 186 pmol pegRNA, and 62 pmol nicking sgRNA were used. Data and error bars represent the mean and standard deviation of three or more independent biological replicates. The n values for PE2 and PE3 in figures c-e are indicated alongside each sample type. Data analysis used One-way ANOVA with Tukey's multiple comparisons test; NS, not significant; *p < 0.05; **p < 0.01; ***p < 0.001. *pegRNA*: ligated pegRNA by a 32 nt synthetic acceptor RNA with 5′ end modifications and synthetic donor RNA with 3′ end modifications. *epegRNA: ligated epegRNA by a 32 nt synthetic acceptor RNA with 5′ modifications and IVT-generated donor RNA containing evopreQ1. *epegRNA*: ligated epegRNA by a 91 nt synthetic acceptor RNA with 5′ end modifications and synthetic donor RNA with 3′ end modifications at evopreQ1 sequence.
