In vivo cytidine base editing of hepatocytes without detectable off-target mutations in RNA and DNA
Abstract
Base editors are RNA-programmable deaminases that enable precise single-base conversions in genomic DNA. However, off-target activity is a concern in the potential use of base editors to treat genetic diseases. Here, we report unbiased analyses of transcriptome-wide and genome-wide off-target modifications effected by cytidine base editors in the liver of mice with phenylketonuria. The intravenous delivery of intein-split cytidine base editors by dual adeno-associated viruses led to the repair of the disease-causing mutation without generating off-target mutations in the RNA and DNA of the hepatocytes. Moreover, the transient expression of a cytidine base editor mRNA and a relevant single-guide RNA intravenously delivered by lipid nanoparticles led to ~21% on-target editing and to the reversal of the disease phenotype; there were also no detectable transcriptome-wide and genome-wide off-target edits. Our findings support the feasibility of therapeutic cytidine base editing to treat genetic liver diseases. Show more
Publication status
publishedExternal links
Journal / series
Nature Biomedical EngineeringVolume
Pages / Article No.
Publisher
NatureSubject
Genetic engineering; Targeted gene repairOrganisational unit
03739 - Stoffel, Markus / Stoffel, Markus
09492 - Schwank, Gerald (ehemalig) / Schwank, Gerald (former)
02207 - Functional Genomics Center Zurich / Functional Genomics Center Zurich
03760 - Hall, Jonathan / Hall, Jonathan
Funding
185293 - Establishment of in vivo CRISPR-Cas base editor approaches to treat monogenetic liver diseases (SNF)
169612 - Chemical Approaches to Functionalize Human microRNAs (SNF)
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