A dual-deaminase CRISPR base editor enables concurrent adenine and cytosine editing

Julian Grünewald; Ronghao Zhou; Caleb A Lareau; Sara P Garcia; Sowmya Iyer; Bret R Miller; Lukas M Langner; Jonathan Y Hsu; Martin J Aryee; J Keith Joung

doi:10.1038/s41587-020-0535-y

A dual-deaminase CRISPR base editor enables concurrent adenine and cytosine editing

Nat Biotechnol. 2020 Jul;38(7):861-864. doi: 10.1038/s41587-020-0535-y. Epub 2020 Jun 1.

Authors

Julian Grünewald^#^{1

2

3}, Ronghao Zhou^#^{1

2}, Caleb A Lareau^{1

4}, Sara P Garcia¹, Sowmya Iyer¹, Bret R Miller^{1

2}, Lukas M Langner^{1

2}, Jonathan Y Hsu^{1

2

5}, Martin J Aryee^{1

2

3

4}, J Keith Joung^{6

7

8}

Affiliations

¹ Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA, USA.
² Center for Cancer Research and Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, MA, USA.
³ Department of Pathology, Harvard Medical School, Boston, MA, USA.
⁴ Department of Biostatistics, Harvard T.H. Chan School of Public Health, Boston, MA, USA.
⁵ Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁶ Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA, USA. jjoung@mgh.harvard.edu.
⁷ Center for Cancer Research and Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, MA, USA. jjoung@mgh.harvard.edu.
⁸ Department of Pathology, Harvard Medical School, Boston, MA, USA. jjoung@mgh.harvard.edu.

^# Contributed equally.

Abstract

Existing adenine and cytosine base editors induce only a single type of modification, limiting the range of DNA alterations that can be created. Here we describe a CRISPR-Cas9-based synchronous programmable adenine and cytosine editor (SPACE) that can concurrently introduce A-to-G and C-to-T substitutions with minimal RNA off-target edits. SPACE expands the range of possible DNA sequence alterations, broadening the research applications of CRISPR base editors.

Publication types

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

MeSH terms

Adenine / chemistry
CRISPR-Associated Protein 9 / genetics*
CRISPR-Cas Systems / genetics*
Cytosine / chemistry
Cytosine Deaminase / genetics*
Gene Editing*
HEK293 Cells
Humans
Mutation / genetics
RNA / genetics

Substances

RNA
Cytosine
CRISPR-Associated Protein 9
Cytosine Deaminase
Adenine

Abstract

Publication types

MeSH terms

Substances

Grants and funding