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

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Description

<jats:title>Abstract</jats:title><jats:p>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 <jats:underline>vi</jats:underline>rus-based guide RNA (gRNA) delivery system for CRISPR/Cas9 mediated plant <jats:underline>g</jats:underline>enome <jats:underline>e</jats:underline>diting (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 <jats:italic>NbPDS3</jats:italic> and <jats:italic>NbIspH</jats:italic> genes in non-inoculated leaves because CaLCuV can infect plants systemically. Moreover, VIGE of <jats:italic>NbPDS3</jats:italic> and <jats:italic>NbIspH</jats:italic> in newly developed leaves caused photo-bleached phenotype. These results demonstrate that geminivirus-based VIGE could be a powerful tool in plant genome editing.</jats:p>

Journal

  • Scientific Reports

    Scientific Reports 5 (1), 14926-, 2015-10-09

    Springer Science and Business Media LLC

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