DNA capture by a CRISPR-Cas9–guided adenine base editor-Reference-Cited by-同舟云学术

DNA capture by a CRISPR-Cas9–guided adenine base editor

Published:2020-07-31 Issue:6503 Volume:369 Page:566-571
ISSN:0036-8075
Container-title:Science
language:en
Short-container-title:Science

Author:

Lapinaite Audrone¹^ORCID,Knott Gavin J.¹²^ORCID,Palumbo Cody M.³^ORCID,Lin-Shiao Enrique¹^ORCID,Richter Michelle F.⁴⁵⁶^ORCID,Zhao Kevin T.⁴⁵⁶^ORCID,Beal Peter A.³^ORCID,Liu David R.⁴⁵⁶^ORCID,Doudna Jennifer A.¹⁷⁸⁹¹⁰^ORCID

Affiliation:

1. Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.

2. Monash Biomedicine Discovery Institute, Department of Biochemistry & Molecular Biology, Monash University, Victoria 3800, Australia.

3. Department of Chemistry, University of California, Davis, CA 95616, USA.

4. Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of Harvard and MIT, Cambridge, MA 02141, USA.

5. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.

6. Howard Hughes Medical Institute, Harvard University, Cambridge, MA 02138, USA.

7. Innovative Genomics Institute, University of California, Berkeley, CA 94720, USA.

8. MBIB Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.

9. Department of Chemistry, University of California, Berkeley, CA 94720, USA.

10. Howard Hughes Medical Institute, University of California, Berkeley, CA 94720, USA.

Abstract

Secrets of a fast base editor CRISPR-Cas9 base editors comprise RNA-guided Cas proteins fused to an enzyme that can deaminate a DNA nucleoside. No natural enzyme deaminates adenine in DNA, and so a breakthrough came when a natural transfer RNA deaminase was fused to Cas9 and evolved to give an adenine base editor (ABE) that works on DNA. Further evolution provided the enzyme ABE8e, which catalyzes deamination more than 1000 times faster than early ABEs. Lapinaite et al. now present a 3.2-angstrom resolution structure of ABE8e bound to DNA in which the target adenine is replaced with an analog designed to trap the catalytic conformation. The structure, together with kinetic data comparing ABE8e to earlier ABEs, explains how ABE8e edits DNA bases and could inform future base-editor design. Science , this issue p. 566

Funder

National Science Foundation

National Institutes of Health

Howard Hughes Medical Institute

W.M. Keck Foundation

National Multiple Sclerosis Society

Paul G. Allen Family Foundation

St. Jude Medical Foundation

Bill and Melinda Gates Foundation

Publisher

American Association for the Advancement of Science (AAAS)

Subject