Data from: Improved specificity and efficiency of in vivo adenine base editing therapies with hybrid guide RNAs

Phenylketonuria (PKU), pseudoxanthoma elasticum (PXE), and hereditary
tyrosinemia type 1 (HT1) are autosomal recessive disorders linked to the
PAH, ABCC6, and FAH and HPD genes, respectively. Here we evaluated the
off-target editing profiles of clinical lead guide RNAs (gRNAs) that, when
combined with adenine base editors (ABEs), correct the
recurrent PAH P281L variant, PAH R408W variant,
or ABCC6 R1164X variant or disrupt either of two sites
in the HPD gene (a modifier gene of HT1) in human
hepatocytes. To mitigate off-target mutagenesis, we systematically
screened hybrid gRNAs with DNA nucleotide substitutions. Comprehensive and
variant-aware specificity profiling of these hybrid gRNAs revealed
dramatically reduced off-target editing and reduced bystander editing in
cells. In humanized PAH P281L and ABCC6 R1164X
mouse models of PKU and PXE, we showed that when formulated in lipid
nanoparticles (LNPs) with ABE mRNA, selected hybrid gRNAs reverted disease
phenotypes, reduced off-target editing, increased on-target editing, and
reduced bystander editing in vivo. These studies highlight the
utility of hybrid gRNAs to improve the safety and efficiency of adenine
base editing therapies. We performed ONE‑seq in this manuscript, and here
is the original dataset for all the ONE‑seq experiments conducted for this
study.