Knocking Out MicroRNA Genes in Rice with CRISPR-Cas9

Jianping Zhou; Zhaohui Zhong; Hongqiao Chen; Qian Li; Xuelian Zheng; Yiping Qi; Yong Zhang

doi:10.1007/978-1-4939-8991-1_9

Knocking Out MicroRNA Genes in Rice with CRISPR-Cas9

Methods Mol Biol. 2019:1917:109-119. doi: 10.1007/978-1-4939-8991-1_9.

Authors

Jianping Zhou¹, Zhaohui Zhong¹, Hongqiao Chen¹, Qian Li¹, Xuelian Zheng¹, Yiping Qi^{2

3}, Yong Zhang⁴

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 Plant Science and Landscape Architecture, University of Maryland, College Park, MD, USA.
³ Institute for Bioscience and Biotechnology Research, University of Maryland, Rockville, MD, USA.
⁴ 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: 30610632
DOI: 10.1007/978-1-4939-8991-1_9

Abstract

MicroRNAs (miRNAs) are small noncoding RNAs that play important roles in plant development and stress responses. Loss-of-function analysis of miRNA genes has been traditionally challenging due to lack of appropriate knockout tools. In this chapter, we describe a method of using CRISPR-Cas9 for knocking out microRNA genes in rice by Agrobacterium-mediated transformation. We also demonstrate single-strand conformation polymorphism (SSCP) as an effective genotyping method for screening CRISPR-Cas9-induced mutations.

Keywords: CRISPR-Cas9; Genome editing; MicroRNAs; Rice; SSCP.

Publication types

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

MeSH terms

CRISPR-Cas Systems / genetics
CRISPR-Cas Systems / physiology
Genome, Plant / genetics*
MicroRNAs / genetics*
Oryza / genetics*

Substances

MicroRNAs