Targeted Base Editing with CRISPR-Deaminase in Tomato

Zenpei Shimatani; Tohru Ariizumi; Ushio Fujikura; Akihiko Kondo; Hiroshi Ezura; Keiji Nishida

doi:10.1007/978-1-4939-8991-1_22

Targeted Base Editing with CRISPR-Deaminase in Tomato

Methods Mol Biol. 2019:1917:297-307. doi: 10.1007/978-1-4939-8991-1_22.

Authors

Zenpei Shimatani¹, Tohru Ariizumi², Ushio Fujikura¹, Akihiko Kondo^{1

3}, Hiroshi Ezura², Keiji Nishida⁴

Affiliations

¹ Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Hyogo, Japan.
² Faculty of Life and Environmental Sciences, Gene Research Center, University of Tsukuba, Tsukuba, Ibaraki, Japan.
³ Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Kobe, Hyogo, Japan.
⁴ Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Hyogo, Japan. keiji_nishida@people.kobe-u.ac.jp.

PMID: 30610645
DOI: 10.1007/978-1-4939-8991-1_22

Abstract

The Target-AID system, consisting of a complex of cytidine deaminase and deficient CRISPR/Cas9, enables highly specific genomic nucleotide substitutions without the need for template DNA. The Cas9-fused cytidine deaminase is guided by sgRNAs and catalyzes the conversion of cytosine to uracil. The resulting U-G DNA mismatches trigger nucleotide substitutions (C to T or G to A) through DNA replication and repair pathways. Target-AID also retains the benefits of conventional CRISPR/Cas9 including robustness in various organisms, high targeting efficiency, and multiplex simultaneous gene editing. Our research group recently developed plant-optimized Target-AID system and demonstrated targeted base editing in tomato and rice. In this chapter, we introduce methods for Target-AID application in tomato.

Keywords: Activation-induced cytidine deaminase (AID); CRISPR/Cas9; Target-AID; Targeted nucleotide substitution; Tomato.

Publication types

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

MeSH terms

CRISPR-Cas Systems / genetics*
Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
Cytidine Deaminase / genetics
Gene Editing / methods*
Solanum lycopersicum / genetics*

Substances

Cytidine Deaminase