Base editors for simultaneous introduction of C-to-T and A-to-G mutations

Rina C Sakata; Soh Ishiguro; Hideto Mori; Mamoru Tanaka; Kenji Tatsuno; Hiroki Ueda; Shogo Yamamoto; Motoaki Seki; Nanami Masuyama; Keiji Nishida; Hiroshi Nishimasu; Kazuharu Arakawa; Akihiko Kondo; Osamu Nureki; Masaru Tomita; Hiroyuki Aburatani; Nozomu Yachie

doi:10.1038/s41587-020-0509-0

Base editors for simultaneous introduction of C-to-T and A-to-G mutations

Nat Biotechnol. 2020 Jul;38(7):865-869. doi: 10.1038/s41587-020-0509-0. Epub 2020 Jun 2.

Authors

Rina C Sakata^#^{1

2}, Soh Ishiguro^#^{1

3

4}, Hideto Mori^#^{1

3

4}, Mamoru Tanaka¹, Kenji Tatsuno⁵, Hiroki Ueda⁶, Shogo Yamamoto⁵, Motoaki Seki¹, Nanami Masuyama^{1

3

4}, Keiji Nishida^{7

8}, Hiroshi Nishimasu⁹, Kazuharu Arakawa^{3

4

10}, Akihiko Kondo^{7

8

11}, Osamu Nureki⁹, Masaru Tomita^{1

3

4

10}, Hiroyuki Aburatani⁵, Nozomu Yachie^{12

13

14

15

16

17}

Affiliations

¹ Synthetic Biology Division, Research Center for Advanced Science and Technology, University of Tokyo, Tokyo, Japan.
² College of Arts and Sciences, University of Tokyo, Tokyo, Japan.
³ Institute for Advanced Biosciences, Keio University, Tsuruoka, Japan.
⁴ Graduate School of Media and Governance, Keio University, Fujisawa, Japan.
⁵ Genome Science Division, Research Center for Advanced Science and Technology, University of Tokyo, Tokyo, Japan.
⁶ Biological Data Science Division, Research Center for Advanced Science and Technology, University of Tokyo, Tokyo, Japan.
⁷ Engineering Biology Research Center, Kobe University, Kobe, Japan.
⁸ Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan.
⁹ Department of Biological Sciences, School of Science, University of Tokyo, Tokyo, Japan.
¹⁰ Faculty of Environment and Information Studies, Keio University, Fujisawa, Japan.
¹¹ Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Kobe, Japan.
¹² Synthetic Biology Division, Research Center for Advanced Science and Technology, University of Tokyo, Tokyo, Japan. yachie@synbiol.rcast.u-tokyo.ac.jp.
¹³ College of Arts and Sciences, University of Tokyo, Tokyo, Japan. yachie@synbiol.rcast.u-tokyo.ac.jp.
¹⁴ Institute for Advanced Biosciences, Keio University, Tsuruoka, Japan. yachie@synbiol.rcast.u-tokyo.ac.jp.
¹⁵ Graduate School of Media and Governance, Keio University, Fujisawa, Japan. yachie@synbiol.rcast.u-tokyo.ac.jp.
¹⁶ Department of Biological Sciences, School of Science, University of Tokyo, Tokyo, Japan. yachie@synbiol.rcast.u-tokyo.ac.jp.
¹⁷ PRESTO, Japan Science and Technology Agency (JST), Tokyo, Japan. yachie@synbiol.rcast.u-tokyo.ac.jp.

^# Contributed equally.

PMID: 32483365
DOI: 10.1038/s41587-020-0509-0

Abstract

We describe base editors that combine both cytosine and adenine base-editing functions. A codon-optimized fusion of the cytosine deaminase PmCDA1, the adenosine deaminase TadA and a Cas9 nickase (Target-ACEmax) showed a high median simultaneous C-to-T and A-to-G editing activity at 47 genomic targets. On-target as well as DNA and RNA off-target activities of Target-ACEmax were similar to those of existing single-function base editors.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenine / metabolism
Adenosine Deaminase / genetics
CRISPR-Associated Protein 9 / genetics*
CRISPR-Cas Systems / genetics*
Cytosine / metabolism
DNA / genetics*
Deoxyribonuclease I / genetics
Gene Editing*
Genome, Human / genetics
Glycoproteins / genetics
Guanine / metabolism
HEK293 Cells
Humans
Mutation / genetics
Nuclear Proteins / genetics
RNA / genetics

Substances

CDAN1 protein, human
Glycoproteins
Nuclear Proteins
Guanine
RNA
Cytosine
DNA
CRISPR-Associated Protein 9
Deoxyribonuclease I
Adenosine Deaminase
Adenine

Associated data

figshare/10.6084/m9.figshare.12016785.v1