Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion

Zenpei Shimatani; Sachiko Kashojiya; Mariko Takayama; Rie Terada; Takayuki Arazoe; Hisaki Ishii; Hiroshi Teramura; Tsuyoshi Yamamoto; Hiroki Komatsu; Kenji Miura; Hiroshi Ezura; Keiji Nishida; Tohru Ariizumi; Akihiko Kondo

doi:10.1038/nbt.3833

Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion

Nat Biotechnol. 2017 May;35(5):441-443. doi: 10.1038/nbt.3833. Epub 2017 Mar 27.

Authors

Affiliations

¹ Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan.
² Faculty of Life and Environmental Sciences, University of Tsukuba, Gene Research Center, Tsukuba, Japan.
³ Laboratory of Genetics and Breeding Science, Faculty of Agriculture, Meijo University, Nagoya, Japan.
⁴ Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Kobe, Japan.

PMID: 28346401
DOI: 10.1038/nbt.3833

Abstract

We applied a fusion of CRISPR-Cas9 and activation-induced cytidine deaminase (Target-AID) for point mutagenesis at genomic regions specified by single guide RNAs (sgRNAs) in two crop plants. In rice, we induced multiple herbicide-resistance point mutations by multiplexed editing using herbicide selection, while in tomato we generated marker-free plants with homozygous heritable DNA substitutions, demonstrating the feasibility of base editing for crop improvement.

Publication types

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

MeSH terms

Base Pairing / genetics
CRISPR-Associated Proteins / genetics*
Clustered Regularly Interspaced Short Palindromic Repeats / genetics
Cytidine Deaminase / genetics
DNA, Plant / genetics*
Gene Editing / methods*
Genes, Plant / genetics
Mutagenesis, Site-Directed / methods*
Oryza / genetics*
Plants, Genetically Modified / genetics
Point Mutation / genetics
Recombinant Fusion Proteins / genetics
Solanum lycopersicum / genetics*

Substances

CRISPR-Associated Proteins
DNA, Plant
Recombinant Fusion Proteins
Cytidine Deaminase