A geminivirus-based guide RNA delivery system for CRISPR/Cas9 mediated plant genome editing

Journal Article / 2015

Yin, Kangquan; Han, Ting; Liu, Guang; Chen, Tianyuan; Wang, Ying; Yu, Alice Yunzi L.; Liu, Yule

CRISPR/Cas has emerged as potent genome editing technology and has successfully been applied in many organisms, including several plant species. However, delivery of genome editing reagents remains a challenge in plants. Here, we report a virus-based guide RNA (gRNA) delivery system for CRISPR/Cas9 mediated plant genome editing (VIGE) that can be used to precisely target genome locations and cause mutations. VIGE is performed by using a modified Cabbage Leaf Curl virus (CaLCuV) vector to express gRNAs in stable transgenic plants expressing Cas9. DNA sequencing confirmed VIGE of endogenous NbPDS3 and NbIspH genes in non-inoculated leaves because CaLCuV can infect plants systemically. Moreover, VIGE of NbPDS3 and NbIspH in newly developed leaves caused photo-bleached phenotype. These results demonstrate that geminivirus-based VIGE could be a powerful tool in plant genome editing.

CRISPR-Cas Systems/genetics; DNA, Plant/genetics/metabolism; Geminiviridae/genetics; Genetic Engineering/methods; Genetic Vectors/genetics; Genome, Plant/genetics; Models, Genetic; Plant Leaves/genetics/virology; Plants, Genetically Modified; Plants/genetics/virology; Reproducibility of Results; RNA, Guide/genetics; Tobacco/genetics/virology

Scientific reports

14926

5

10.1038/srep14926

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