A Single Transcript CRISPR-Cas9 System for Multiplex Genome Editing in Plants

Xu Tang; Zhaohui Zhong; Qiurong Ren; Binglin Liu; Yong Zhang

doi:10.1007/978-1-4939-8991-1_6

A Single Transcript CRISPR-Cas9 System for Multiplex Genome Editing in Plants

Methods Mol Biol. 2019:1917:75-82. doi: 10.1007/978-1-4939-8991-1_6.

Authors

Xu Tang¹, Zhaohui Zhong¹, Qiurong Ren¹, Binglin Liu¹, Yong Zhang²

Affiliations

¹ Department of Biotechnology, School of Life Science and Technology, Center for Informational Biology, University of Electronic Science and Technology of China, Chengdu, China.
² Department of Biotechnology, School of Life Science and Technology, Center for Informational Biology, University of Electronic Science and Technology of China, Chengdu, China. zhangyong916@uestc.edu.cn.

PMID: 30610629
DOI: 10.1007/978-1-4939-8991-1_6

Abstract

The CRISPR-Cas9 system has been widely adopted in genome editing. By changing the 20 bp guide sequence, it can easily edit any sequence adjacent to a protospacer adjacent motif (PAM) in a genome. Multiplex genome editing could be accomplished with simultaneous expression of multiple single-guide RNAs (sgRNA). Given that sgRNAs are expressed by Pol III promoters, multiplex genome editing is conventionally done by assembly of multiple complete sgRNA expression cassettes together, which can be a challenge in vector construction. Here, we described a multiplex genome editing system based on a single transcript unit CRISPR-Cas9 (STU CRISPR-Cas9) expression system driven by a single Pol II promoter. It represents a novel approach for multiplex genome editing.

Keywords: CRISPR-Cas9; Multiplex genome editing; STU.

Publication types

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

MeSH terms

CRISPR-Cas Systems / genetics*
Gene Editing / methods*
Genome, Plant / genetics*
Plants / genetics*
Promoter Regions, Genetic / genetics
RNA Polymerase II / genetics
RNA, Guide, CRISPR-Cas Systems / genetics

Substances

RNA, Guide, CRISPR-Cas Systems
RNA Polymerase II