Fig. 2

Desired edits and unwanted byproducts in the two transformations with pZ1WS. a Desired edits and unwanted byproducts at the two target sites in the first transformation. Only partial sgRNA sequences are shown. The homologous sgRNA-rtT sequences shared by all the aligned sequences are shaded in yellow, and the mutated nucleotides are indicated by red letters. The number of cloned PCR fragments harboring the same edits is indicated in parentheses, and a vertical line indicates the same type of byproduct. For convenience, the byproducts derived from the two mechanisms were assorted into pegRNA scaffold-derived byproduct category. b Summary of the desired edits and byproducts from the first transformation according to sequences of the cloned PCR fragments. c Prime-editing efficiency achieved in the second transformation. Strong and weak, T0 lines harboring strong and weak peak signals for the edits, respectively, in the sequencing chromatograms. Both, total no. of lines harboring strong and weak edits. d Sequencing chromatograms from a prime-edited line harboring W542L edits and obtained in the second transformation. Double peaks represent heterozygous or chimeric mutations, and an asterisk indicates a mutation induced by PE. The first mutation in the S621 target is the pegRNA scaffold-derived byproducts. e Desired edits and unwanted byproducts at the two target sites in the second transformation. f Summary of the desired edits and byproducts in the second transformation