Extended Data Fig. 2: Split-PE, SunTag-PE3 and MS2-PE3 tested in an mCherry reporter line and an endogenous locus.
From: A split prime editor with untethered reverse transcriptase and circular RNA template
a, A diagram of the mCherry reporter line that functions by converting a premature stop codon. b, Sequences of RTT and PBS, non-cognate (PBS + RTT), non-cognate PBS, and non-cognate RTT for the mCherry reporter line. c, Multiple MS2-pegRNAs tested in mCherry reporter cell lines. The pegRNA with MS2 on the repeat/anti-repeat stem-loop (pegRNA-1.1) has the highest editing efficiency (higher even than that of the original PE3) in this mCherry reporter line (n = 2). Therefore, the pegRNA1.1-Cas9H840A-MCP-RT system was designated as MS2-PE3. d, SunTag-PE3 and PE3-SunTag were tested in the mCherry reporter cell line. Two-tailed unpaired Student’s t-test: *P < 0.05 (n = 3). e, Sanger sequencing and EditR quantification of PE3, Split PE, SunTag-PE3 and MS2-PE3 by installing “CTT” at HEK3 sites in HEK293T cells. All plasmids were transfected at the same molar ratio. Genomic DNAs were isolated 72 h post transfection. f, Dose dependence of the RT-encoding plasmid. One microgram of H840A plasmid was co-transfected with plasmids encoding additional sPE components [pegRNA (0.3 µg), nicking sgRNA (0.1 µg), and RT (0.01-2 µg)] per well in a 12-well plate (n = 2). Data and error bars indicate the mean and standard deviation of two or three independent biological replicates, as indicated.
