Optimization of CRISPR-Cas9 System in

Overview

Journal iScience

Publisher Cell Press

Date 2024 Feb 16

PMID 38361623

Authors

Xueqi Li

Fanqi Bu

Lishan Wang

Cholmin Kim

Wanjie Xue

Man Zhang

Saneyuki Kawabata

Qingzhu Zhang

Yuhua Li

Yang Zhang

Affiliations

Soon will be listed here.

Abstract

The optimization of the CRISPR-Cas9 system for enhancing editing efficiency holds significant value in scientific research. In this study, we optimized single guide RNA and promoters of the CRISPR-Cas9 vector and established an efficient protoplast isolation and transient transformation system in , and we successfully applied the modified CRISPR-Cas9 system to detect editing efficiency of the gene. The activity of the promoter in protoplasts was approximately three times higher than that of the promoter. This promoter, along with the promoter, was applied in the CRISPR-Cas9 cassette, the modified CRISPR-Cas9 vectors that editing efficiency was 37.7%, which was 30.3% higher than that of the control, and the types of mutation are base substitutions, small fragment deletions and insertions. Finally we obtained an efficient gene editing vector for . This project provides an important technical platform for the study of gene function in .

Citing Articles

Establishing a CRISPR/Cas9 genome editing framework in pigeonpea (Cajanus cajan L.) by targeting phytoene desaturase (PDS) gene disruption.

Senthil K, Rathinam M, Parashar M, Dokka N, Tyagi S, Mathur V J Genet Eng Biotechnol. 2025; 23(1):100465.

PMID: 40074438 PMC: 11847732. DOI: 10.1016/j.jgeb.2025.100465.

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