Genome editing in rice and wheat using the CRISPR/Cas system

Area of Science:

• Plant biotechnology
• Molecular biology
• Genomics

Background:

• Zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) are established genome editing tools.
• The clustered regularly interspersed short palindromic repeats (CRISPR)/Cas system offers a versatile alternative for genome engineering.

Purpose of the Study:

• To describe a protocol for CRISPR/Cas-mediated gene targeting in rice and wheat.
• To facilitate sequence-specific mutagenesis and gene targeting using the CRISPR/Cas system.

Main Methods:

• Selection of target sites for single-guide RNA (sgRNA).
• Design, construction, and verification of sgRNAs.
• CRISPR/Cas-mediated mutagenesis and gene targeting in rice and wheat protoplasts and plants.

Main Results:

• The CRISPR/Cas system allows for rapid gene targeting in protoplasts within 1-2 weeks.
• Mutated rice plants were generated within 13-17 weeks.
• The protocol simplifies the process of targeting specific genes in plants.

Conclusions:

• The CRISPR/Cas system is a powerful and accessible tool for plant genome engineering.
• This protocol streamlines gene targeting in rice and wheat, aiding in the development of new plant traits.