Extended Data Fig. 5: Generalization of benefits of hybrid gRNAs to additional loci.
From: Improved specificity and efficiency of in vivo adenine base editing therapies with hybrid guide RNAs
a, Spacer sequences of four additional sets of standard and hybrid gRNAs. ‘rN’ = ribonucleotide; ‘dN’ = deoxyribonucleotide; ‘m’ = 2’-O-methylation; * = phosphorothioate linkage. DNA substitutions are in red bold and underlined. The target variant adenine bases are in blue bold and underlined. b, A-to-G editing of on-target sites in lentivirus-transduced HuH-7 cells with the variants to be corrected, treated with standard or hybrid gRNA in combination with mRNA (n = 3 biological replicates per condition). For the CPS1-1 gRNAs, essentially all bystander editing resulted in synonymous variants, and so no unwanted bystander editing is shown. Means are shown. c, Number of sites with ONE-seq score > 0.01 for each indicated standard or hybrid gRNA with adenine base editor protein. ONE-seq for the PAH5_hyb16 gRNA was deferred because, as shown in b, the desired on-target editing with PAH5_hyb16 was less than that with the standard PAH5 gRNA.
