In vivo cytidine base editing of hepatocytes without detectable off-target mutations in RNA and DNA
- 25 January 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Biomedical Engineering
- Vol. 5 (2), 179-189
- https://doi.org/10.1038/s41551-020-00671-z
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.Keywords
Funding Information
- Novartis Stiftung für Medizinisch-Biologische Forschung (19A004)
- Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (205 321_169 612, 310030_185293)
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