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. Author manuscript; available in PMC: 2023 Jun 6.
Published in final edited form as: Nat Biotechnol. 2022 Apr 11;40(6):874–884. doi: 10.1038/s41587-022-01276-4

Extended Data Fig. 2 |. Generation of libraries C and A.

Extended Data Fig. 2 |

a, The process of selecting sgRNA-target pairs for the generation of libraries C and A. SNVs, single nucleotide variants; sgRNA, single guide RNA. b, Generation of lentiviral libraries of sgRNA-encoding and target sequence pairs with unique molecular identifiers (UMIs). Oligonucleotides containing a 20-nt guide sequence, and the corresponding target sequence were synthesized and cloned into the pLenti-gRNA-puro vector to create plasmid library 1. The plasmids were then digested with BsmBI restriction enzyme and ligated with fragments containing the sgRNA scaffold sequences and UMIs to create plasmid library 2. Lentiviral libraries generated from plasmid library 2 were then transduced into cells expressing cytosine base editor (CBE) or adenine base editor (ABE) in a doxycycline-inducible manner.