Extended Data Fig. 5: Prime editing at on target site and two off target sites in HEK293T cells.
From: Genome-wide profiling of prime editor off-target sites in vitro and in vivo using PE-tag
a, (left) Comparison of precise editing efficiency for nucleotide substitution, targeted 1-bp deletion, and 1-bp insertion with PE2 at HEK4 (ON) target site and (right) indel rates at two off-target sites (OT-1 and OT-3) in HEK293T cells after co-transfecting pegRNA and PE2 expression plasmids. pegRNA sequence composition and type of sequence modification encoded is indicated in the legend, where the terminal numbers indicate the different HA lengths within the RTT. Frequencies of precise editing or indel rates were quantified by deep sequencing from PCR amplicons spanning each locus. Mock on target site editing represents all indels. Results were obtained from three independent experiments and presented as mean ± SD. *P < 0.05, ** P < 0.01 and *** P < 0.001 by unpaired, two-tailed Student’s t-test. To adjust for multiple comparisons, p-values were adjusted using the Benjamini-Hochberg (BH) method. b, indel rates for nucleotide substitution, targeted 1-bp deletion, and 1-bp insertion pegRNAs with PE2 at 6 additional potential off-target sites identified by PE-tag for HEK4 locus pegRNA in HEK293T cells. pegRNA sequence composition and type of sequence modification encoded is indicated in the legend, where the terminal numbers indicate the different HA lengths within the RTT. Frequencies of precise editing were quantified by deep sequencing from PCR amplicons spanning each locus. Results were obtained from three independent experiments and presented as mean ± SD. *P < 0.05, ** P < 0.01 and *** P < 0.001 by unpaired, two-tailed Student’s t-test. To adjust for multiple comparisons, p-values were adjusted using the Benjamini-Hochberg (BH) method.
