Structure and DNA sequences of synthetic transcription cassettes within plasmids expressing unmodified and modified pegRNAs.

(A) Schematic diagrams of synthetic transcription cassettes expressing pegRNAs and epegRNAs (tmpknot and tevopreQ1) targeted to mut-1, mut-2, and non-target controls (V.C.). Genetic components depicted with color squares are connected without intervening sequences in the plasmids indicated on the left. (B) DNA sequences of the genetic components depicted in (A). Nucleotides indicated by red and blue bold letters represent mutant and wild-type sequences at the edited positions, respectively. Underlining indicates restriction enzyme recognition sites. The lowercase letter “g” indicates a guanine appended to the 5′ end of mut-2 spacer for enhanced transcription. epegRNA, engineered prime editing guide RNA; RTT, reverse transcription template; PBS, primer binding site. (PDF)