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. Author manuscript; available in PMC: 2022 Jun 9.
Published in final edited form as: Nat Biotechnol. 2021 Dec 9;40(5):731–740. doi: 10.1038/s41587-021-01133-w

Extended Data Fig. 2. Recoding of PAH exon sequences in HEK293T cells via twinPE.

Extended Data Fig. 2

Screen of pegRNA pairs targeting PAH for recoding of (a) exons 2, 4 and 5, (b) exon 7, and (c) exons 9, 10, 11, and 12. RT templates of pegRNAs encoded partially recoded exonic sequence to optimize orthogonality to the endogenous gene sequence. For each spacer pair, nine pegRNA combinations were tested using three PBS variants for each spacer in a three-by-three matrix, with RT templates encoding the recoded exonic sequence, which was held constant for given spacer pairs. Sequences of pegRNAs are listed in Supplementary Table 1. Sequences of recoded exonic sequences are listed in Supplementary Table 4. Values in (a), (b) and exon 9 in (c) reflect single biological replicates. Values for exons 10, 11 and 12 in (c) reflect the mean of three independent biological replicates.