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Adenine base editing in an adult mouse model of tyrosinaemia

Nature Biomedical Engineering volume 4, pages 125–130 (2020)Cite this article

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Abstract

In contrast to traditional CRISPR–Cas9 homology-directed repair, base editing can correct point mutations without supplying a DNA-repair template. Here we show in a mouse model of tyrosinaemia that hydrodynamic tail-vein injection of plasmid DNA encoding the adenine base editor (ABE) and a single-guide RNA (sgRNA) can correct an A>G splice-site mutation. ABE treatment partially restored splicing, generated fumarylacetoacetate hydrolase (FAH)-positive hepatocytes in the liver, and rescued weight loss in mice. We also generated FAH+ hepatocytes in the liver via lipid-nanoparticle-mediated delivery of a chemically modified sgRNA and an mRNA of a codon-optimized base editor that displayed higher base-editing efficiency than the standard ABEs. Our findings suggest that adenine base editing can be used for the correction of genetic diseases in adult animals.

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Fig. 1: Adenine base editing rescues liver disease phenotype in a mouse model of tyrosinaemia.
Fig. 2: Adenine base editing partially corrects the Fah mutation in mouse liver.
Fig. 3: Optimizing the coding sequence of ABE6.3 and adding an N-terminal nuclear localization sequence improves base editing.
Fig. 4: RA6.3 shows a higher editing efficiency compared to ABE6.3 and Cas9-mediated HDR at two genomic sites in HEK293T cells.
Fig. 5: RA6.3 increases editing efficiency in vivo compared to ABE6.3.

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Data availability

The authors declare that all data supporting the findings of this study are available within the paper and its Supplementary Information. The deep-sequencing data are available from the Sequence Read Archive (SRA) under accession number PRJNA513076 and PRJNA513291.

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Acknowledgements

We thank C. Mello, P. Zamore, S. Wolfe and E. Sontheimer for discussions; J. Smith for editing the manuscript; M. Grompe (Oregon Health and Science University) for providing the Fah mice; Y. Liu and E. Kittler of the UMass Morphology and Deep Sequencing Cores for support. W.X. was supported by grants from the National Institutes of Health ((NIH) DP2HL137167, P01HL131471 and UG3HL147367), American Cancer Society (129056-RSG-16-093), the Lung Cancer Research Foundation, Hyundai Hope on Wheels, UMass CCTS and ALS Association. This work was supported by DARPA HR0011-17-2-0049; US NIH RM1 HG009490, R01 EB022376, U01 AI142756 and R35 GM118062; and HHMI (to D.R.L.), and R01 CA195787; K22 CA181280 (to L.E.D.). This work was supported in part by the Marble Center for Cancer Nanomedicine and a Cancer Center Support (core) grant P30-CA14051 from the National Cancer Institute. M.R. was supported by the HHMI Hanna H. Gray Fellowship. L.W.K. is an NSF Graduate Research Fellow and was supported NIH Training Grant T32 GM095450. H.Y. was supported by the National Natural Science Foundation of China 31871345, the Young Thousand Talented Program from Wuhan University and startup funding from Wuhan University.

Author information

Author notes

  1. These authors contributed equally: Chun-Qing Song, Tingting Jiang.

Authors and Affiliations

  1. RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA

    Chun-Qing Song, Tingting Jiang, Yueying Cao & Wen Xue

  2. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA

    Michelle Richter, Luke W. Koblan, Jordan L. Doman & David R. Liu

  3. Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA

    Michelle Richter, Luke W. Koblan, Jordan L. Doman & David R. Liu

  4. Broad Institute of MIT and Harvard, Cambridge, MA, USA

    Michelle Richter, Luke W. Koblan, Jordan L. Doman & David R. Liu

  5. David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA

    Luke H. Rhym & Daniel G Anderson

  6. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    Luke H. Rhym & Daniel G Anderson

  7. Sandra and Edward Meyer Cancer Center, Department of Medicine, Weill Cornell Medicine, New York, NY, USA

    Maria Paz Zafra, Emma M. Schatoff & Lukas E. Dow

  8. Weill Cornell/Rockefeller/Sloan Kettering Tri-I MD-PhD program, New York, NY, USA

    Emma M. Schatoff

  9. Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, MA, USA

    Lihua Julie Zhu & Wen Xue

  10. Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, MA, USA

    Lihua Julie Zhu

  11. Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA

    Lihua Julie Zhu & Wen Xue

  12. Medical Research Institute, Wuhan University, Wuhan, China

    Hao Yin

  13. Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, MA, USA

    Wen Xue

Authors
  1. Chun-Qing Song
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Contributions

C.Q.S., T.J., M.R., D.R.L., H.Y. and W.X. designed the study. C.Q.S., T.J., M.R., L.H.R., L.W.K., M.P.Z., E.M.S., J.L.D., Y.C., L.J.Z., L.E.D. and D.G.A. performed experiments and analysed data. C.Q.S., T.J., H.Y. and W.X. wrote the manuscript with comments from all authors.

Corresponding authors

Correspondence to David R. Liu, Hao Yin or Wen Xue.

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Competing interests

D.R.L. is a consultant and co-founder of Editas Medicine, Pairwise Plants and Beam Therapeutics, which are companies that use genome editing. The other authors declare no competing interests.

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Reporting Summary

Supplementary Dataset 1

Guide-seq off-target sites.

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Song, CQ., Jiang, T., Richter, M. et al. Adenine base editing in an adult mouse model of tyrosinaemia. Nat Biomed Eng 4, 125–130 (2020). https://doi.org/10.1038/s41551-019-0357-8

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