Chinese Bulletin of Botany ›› 2021, Vol. 56 ›› Issue (1): 44-49.DOI: 10.11983/CBB20203

• INVITED PROTOCOLS • Previous Articles     Next Articles

CRISPR-based DNA Fragment Deletion in Plants

Xianrong Xie, Dongchang Zeng, Jiantao Tan, Qinlong Zhu, Yaoguang Liu*()   

  1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China
  • Received:2020-12-11 Accepted:2021-02-25 Online:2021-01-01 Published:2021-02-25
  • Contact: Yaoguang Liu

Abstract: CRISPR/Cas9-based genome editing technology has been an important tool to study the gene function and genomic modification. Directed by a guide RNA, Cas9 protein can cleavage the genomic DNA at the target site, and produce mutations, including deletion, insertion, substitution and fragment deletion, by DNA double strand break (DSB) repair mechanism. In this protocol, we introduce the method to use CRISPR/Cas9 system to increase the efficiency of genomic DNA fragment deletion with microhomology-mediated end joining, especially the details in target design and detection of mutant plants.

Key words: genome editing, CRISPR/Cas9, MMEJ, microhomologous, fragment deletion