Generating Mutant for a Stress-Responsive Gene in Rice Using CRISPR-Cas9 System

Chudamani Sharma Prakash; Ramanjulu Sunkar

doi:10.1007/978-1-0716-3973-3_20

Generating Mutant for a Stress-Responsive Gene in Rice Using CRISPR-Cas9 System

Methods Mol Biol. 2024:2832:281-290. doi: 10.1007/978-1-0716-3973-3_20.

Authors

Chudamani Sharma Prakash¹, Ramanjulu Sunkar²

Affiliations

¹ Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK, USA.
² Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK, USA. [email protected].

PMID: 38869803
DOI: 10.1007/978-1-0716-3973-3_20

Abstract

CRISPR/Cas9 system is one of the most often utilized engineering tools for genome editing in many organisms including crop plants and presents great value in both basic and applied research. This is a preferred method because of its relative simplicity, cost-effectiveness, and reliability. The Cas9 nuclease guided by a short single guide RNA (gRNA) can generate double-strand DNA breaks (DSB) at the specific sites in chromosomal DNA. The DSB site is repaired by error-prone repair methods. During repair, some nucleotides are deleted or added at the target site. Here, we present a simplified protocol for generating mutants in gene of interest in rice using CRISPR/Cas9.

Keywords: Agrobacterium; Cas9; Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR); Mutation; Transformation.

MeSH terms

CRISPR-Cas Systems*
DNA Breaks, Double-Stranded
Gene Editing* / methods
Mutation
Oryza* / genetics
Plants, Genetically Modified / genetics
RNA, Guide, CRISPR-Cas Systems* / genetics
Stress, Physiological / genetics

Substances

RNA, Guide, CRISPR-Cas Systems