In vivo cytidine base editing of hepatocytes without detectable off-target mutations in RNA and DNA

Lukas Villiger, Tanja Rothgangl, Dominik Witzigmann, Rurika Oka, Paulo J C Lin, Weihong Qi, Sharan Janjuha, Christian Berk, Femke Ringnalda, Mitchell B Beattie, Markus Stoffel, Beat Thöny, Jonathan Hall, Hubert Rehrauer, Ruben van Boxtel, Ying K Tam, Gerald Schwank

Onderzoeksoutput: Bijdrage aan tijdschriftArtikelpeer review

Samenvatting

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.

Originele taal-2Engels
Pagina's (van-tot)179-189
Aantal pagina's11
TijdschriftNature biomedical engineering
Volume5
Nummer van het tijdschrift2
DOI's
StatusGepubliceerd - feb. 2021

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