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

ABSTRACT: 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.

SUBMITTER: Yin K

PROVIDER: S-EPMC4598821 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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

Yin Kangquan K Han Ting T Liu Guang G Chen Tianyuan T Wang Ying Y Yu Alice Yunzi L AY Liu Yule Y

Scientific reports 20151009

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 ...[more]

PMID: 26450012

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