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

Dataset DOI: [10.5061/dryad.zkh1893pc](10.5061/dryad.zkh1893pc)

## Description of the data and file structure

We performed ONE‑seq in the study titled **“Improved Specificity and Efficiency of In Vivo Adenine Base Editing Therapies with Hybrid Guide RNAs.”** Here is the original dataset for all ONE‑seq experiments conducted for this work.

In each tab, the tab name corresponds to the ONE-seq name. Within each tab:

* **Column A** contains the aligned sequence in each ONE-seq library.
* **Column B** shows the chromosome location of the sequence in Column A.
* **Column C** indicates the orientation of the sequence in Column A.
* **Column D** provides the mismatch information of the sequence compared to the on-target sequence.
* **Column E** lists the calculated ONE-seq score.

## Code/software

The analysis pipeline described in the paper below was used to process the data....,