Meixia Wang, Fang Yan, Huanbin Zhou

STAR Protocol Volume 3, Issue 4, 16 December 2022, 101865

Highlights

• - Description of high-speed tissue culture of rice
• - Steps to prepare CRISPR plasmids and perform A. tumefaciens-mediated rice transformation
• - Generation of precise nucleotide mutations by an adenine base editor
• - Genotyping of genome-edited plants to identify potential mutations in the target region

Summary

Base editing is a precision genome-editing approach that is widely utilized to generate single-nucleotide variants (SNVs) in genomes. Here, we present a protocol to perform targeted adenine (A)-to-guanine (G) substitution in rice using adenine base editor (ABE). We detail the design of sgRNA, CRISPR plasmid construction, rapid genetic transformation of rice, and genotyping of editing events. This protocol can be applied to cytosine base editing in rice as well.

For complete details on the use and execution of this protocol, please refer to Yan et al. (2021).

See https://www.sciencedirect.com/science/article/pii/S2666166722007456

Figure: Base editing is a precision genome-editing approach that is widely utilized to generate single-nucleotide variants (SNVs) in genomes. Here, we present a protocol to perform targeted adenine (A)-to-guanine (G) substitution in rice using adenine base editor (ABE). We detail the design of sgRNA, CRISPR plasmid construction, rapid genetic transformation of rice, and genotyping of editing events. This protocol can be applied to cytosine base editing in rice as well.