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Cited by 105CRISPR and Genetic EngineeringRNA regulation and diseaseVirus-based gene therapy research

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

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Villiger, Lukas, Rothgangl, Tanja, Witzigmann, Dominik, Oka, Rurika, Lin, Paulo J. C., Qi, Weihong, Janjuha, Sharan, Berk, Christian +9 more·2021·Nature Biomedical Engineering
DOI 10.1038/s41551-020-00671-zPMC PMC7610981
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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.

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  • An optimal weight ratio of base editor mRNA to sgRNA within the lead LNP formulation (3:1) was identified, differing from previous reports of LNPs co-encapsulating base editor mRNA and sgRNA.

    “An optimal weight ratio of base editor mRNA to sgRNA within the lead LNP formulation (3:1) was also identified, differing from previous reports of LNPs co-encapsulating base editor mRNA and sgRNA (16, 40).”

    From Ionizable Lipid Nanoparticles for Therapeutic Base Editing of Congenital Brain Disease (2023) · cites this paper as [40]

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Authors
Villiger, L., Rothgangl, T., Witzigmann, D., Oka, R., Lin, P. J. C., Qi, W., Janjuha, S., Berk, C., Ringnalda, F., Beattie, M. B., Stoffel, M., Thöny, B., Hall, J., Rehrauer, H., van Boxtel, R., Tam, Y. K., & Schwank, G.
Affiliations
  • University of Zurich (CH)
  • University of Basel (CH)
  • Oncode Institute (NL)
  • Acuitas Therapeutics (Canada) (CA)
  • ETH Zurich (CH)
  • University Children's Hospital Zurich (CH)
Corresponding author
Schwank, Gerald
ORCID
Venue
Nature Biomedical Engineering
Publisher
Springer Science and Business Media LLC
Published
2021-01-25
Volume
5
Issue
2
Pages
179-189
ISSN
2157-846X (ISSN-L)
DOI
10.1038/s41551-020-00671-z
PMC
PMC7610981
OpenAlex
W3125206715
Cited by
105
References
8
Citations by year
2026: 16 · 2025: 14 · 2024: 24 · 2023: 25 · 2022: 18 · 2021: 8
FWCI
5.88
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Biomedical Engineering (publishedVersion)
  • PubMed (publishedVersion)
  • Zurich Open Repository and Archive (University of Zurich) (acceptedVersion, other-oa)
  • PubMed Central (acceptedVersion)
  • Zurich Open Repository and Archive (University of Zurich) (submittedVersion, other-oa)
  • Zurich Open Repository and Archive (University of Zurich)
Funders
  • Novartis Stiftung für Medizinisch-Biologische Forschung (19A004)
  • Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (205 321_169 612, 310030_185293)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
Keywords
off-target mutations, phenylketonuria, hepatocytes, adeno-associated virus vectors, lipid nanoparticles, single guide RNA
MeSH terms
Gene Editing, Adenoviridae, Animals, Cytidine, DNA, Genetic Vectors, Humans, Mice, Inbred C57BL, RNA, Hepatocytes, Mice, HEK293 Cells

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