Adenine base editing in an adult mouse model of tyrosinaemia

Chun-Qing Song; Tingting Jiang; Michelle Richter; Luke H Rhym; Luke W Koblan; Maria Paz Zafra; Emma M Schatoff; Jordan L Doman; Yueying Cao; Lukas E Dow; Lihua Julie Zhu; Daniel G Anderson; David R Liu; Hao Yin; Wen Xue

doi:10.1038/s41551-019-0357-8

Adenine base editing in an adult mouse model of tyrosinaemia

Nat Biomed Eng. 2020 Jan;4(1):125-130. doi: 10.1038/s41551-019-0357-8. Epub 2019 Feb 25.

Authors

Chun-Qing Song^#¹, Tingting Jiang^#¹, Michelle Richter^{2

3

4}, Luke H Rhym^{5

6}, Luke W Koblan^{2

3

4}, Maria Paz Zafra⁷, Emma M Schatoff^{7

8}, Jordan L Doman^{2

3

4}, Yueying Cao¹, Lukas E Dow⁷, Lihua Julie Zhu^{9

10

11}, Daniel G Anderson^{5

6}, David R Liu^{12

13

14}, Hao Yin¹⁵, Wen Xue^{16

17

18

19}

Affiliations

¹ RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA.
² Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
³ Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA.
⁴ Broad Institute of MIT and Harvard, Cambridge, MA, USA.
⁵ David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁶ Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁷ Sandra and Edward Meyer Cancer Center, Department of Medicine, Weill Cornell Medicine, New York, NY, USA.
⁸ Weill Cornell/Rockefeller/Sloan Kettering Tri-I MD-PhD program, New York, NY, USA.
⁹ Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, MA, USA.
¹⁰ Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, MA, USA.
¹¹ Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA.
¹² Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA. drliu@fas.harvard.edu.
¹³ Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA. drliu@fas.harvard.edu.
¹⁴ Broad Institute of MIT and Harvard, Cambridge, MA, USA. drliu@fas.harvard.edu.
¹⁵ Medical Research Institute, Wuhan University, Wuhan, China. haoyin@whu.edu.cn.
¹⁶ RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA. Wen.Xue@umassmed.edu.
¹⁷ Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, MA, USA. Wen.Xue@umassmed.edu.
¹⁸ Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA. Wen.Xue@umassmed.edu.
¹⁹ Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, MA, USA. Wen.Xue@umassmed.edu.

^# Contributed equally.

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Adenine / metabolism*
Animals
Disease Models, Animal
Female
Gene Editing / methods*
HEK293 Cells
Hepatocytes / metabolism
Humans
Hydrolases / genetics
Liver / metabolism
Point Mutation
RNA / administration & dosage
Tyrosinemias / genetics*

Substances

RNA
Hydrolases
fumarylacetoacetase
Adenine

Abstract

Publication types

MeSH terms

Substances

Grants and funding