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    Pea early-browning virus -mediated genome editing via the CRISPR/Cas9 system in Nicotiana benthamiana and Arabidopsis

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    Pea early-browning-Manuscript.pdf
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    Type
    Article
    Authors
    Ali, Zahir cc
    Eid, Ayman
    Ali, Shawkat cc
    Mahfouz, Magdy M. cc
    KAUST Department
    Biological and Environmental Science and Engineering (BESE) Division
    Bioscience Program
    Center for Desert Agriculture
    Laboratory for Genome Engineering
    Plant Science
    Plant Science Program
    Date
    2017-10-17
    Online Publication Date
    2017-10-17
    Print Publication Date
    2018-01
    Permanent link to this record
    http://hdl.handle.net/10754/626046
    
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    Abstract
    The clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated (Cas9) system has enabled efficient genome engineering in diverse plant species. However, delivery of genome engineering reagents, such as the single guide RNA (sgRNA), into plant cells remains challenging. Here, we report the engineering of Tobacco rattle virus (TRV) and Pea early browning virus (PEBV) to deliver one or multiple sgRNAs into Nicotiana benthamiana and Arabidopsis thaliana (Col-0) plants that overexpress a nuclear localization signal containing Cas9. Our data showed that TRV and PEBV can deliver sgRNAs into inoculated and systemic leaves, and this resulted in mutagenesis of the targeted genomic loci. Moreover, in N. benthamiana, PEBV-based sgRNA delivery resulted in more targeted mutations than TRV-based delivery. Our data indicate that TRV and PEBV can facilitate plant genome engineering and can be used to produce targeted mutations for functional analysis and other biotechnological applications across diverse plant species.Key message: Delivery of genome engineering reagents into plant cells is challenging and inefficient and this limit the applications of this technology in many plant species. RNA viruses such as TRV and PEBV provide an efficient tool to systemically deliver sgRNAs for targeted genome modification.
    Citation
    Ali Z, Eid A, Ali S, Mahfouz MM (2017) Pea early-browning virus -mediated genome editing via the CRISPR/Cas9 system in Nicotiana benthamiana and Arabidopsis. Virus Research. Available: http://dx.doi.org/10.1016/j.virusres.2017.10.009.
    Sponsors
    We would like to thank member of the laboratory for genome engineering for continuous discussions. We would like to thank Professor Elisabeth Johansen, Danish Institute for food and veterinary research, for providing PEBV-containing binary constructs. This study is supported by King Abdullah University of Science and Technology (KAUST).
    Publisher
    Elsevier BV
    Journal
    Virus Research
    DOI
    10.1016/j.virusres.2017.10.009
    PubMed ID
    29051052
    Additional Links
    http://www.sciencedirect.com/science/article/pii/S0168170217305543
    ae974a485f413a2113503eed53cd6c53
    10.1016/j.virusres.2017.10.009
    Scopus Count
    Collections
    Articles; Biological and Environmental Science and Engineering (BESE) Division; Bioscience Program; Plant Science Program; Center for Desert Agriculture

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