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

Bibliography:

Zahir Ali, Ayman Eid, Shakila Ali, Magdy M. Mahfouz. Pea early-browning virus-mediated genome editing via the CRISPR/Cas9 system in Nicotiana benthamiana and Arabidopsis. Virus Research. https://doi.org/10.1016/j.virusres.2017.10.009

Authors:

Zahir Ali, Ayman Eid, Shakila Ali, Magdy M. Mahfouz

Keywords:

CRISPR/Cas9, Virus-mediated genome engineering, Virus-mediated genome editing, Targeted mutagenesis, Tobacco rattle virus, Pea early browning virus, Arabidopsis, Nicotiana benthamiana

Year:

2017

Abstract:

‚ÄčThe 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.

ISSN:

0168-1702