Pea early-browning virus -mediated genome editing via the CRISPR/Cas9 system in Nicotiana benthamiana and Arabidopsis
KAUST DepartmentBiological and Environmental Sciences and Engineering (BESE) Division
Desert Agriculture Initiative
Plant Science Program
Laboratory for Genome Engineering, Division of Environmental and Biological Sciences and Engineering, 4700 King Abdullah University of Science and Technology, Thuwal, 23955-6900, Saudi Arabia
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AbstractThe 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.
CitationAli 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.
SponsorsWe 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).
- Activity and specificity of TRV-mediated gene editing in plants.
- Authors: Ali Z, Abul-Faraj A, Piatek M, Mahfouz MM
- Issue date: 2015
- Cas9-based genome editing in Arabidopsis and tobacco.
- Authors: Li JF, Zhang D, Sheen J
- Issue date: 2014
- A CRISPR/Cas9 Toolbox for Multiplexed Plant Genome Editing and Transcriptional Regulation.
- Authors: Lowder LG, Zhang D, Baltes NJ, Paul JW 3rd, Tang X, Zheng X, Voytas DF, Hsieh TF, Zhang Y, Qi Y
- Issue date: 2015 Oct
- Targeted plant genome editing via the CRISPR/Cas9 technology.
- Authors: Li JF, Zhang D, Sheen J
- Issue date: 2015
- CRISPR/Cas9-mediated targeted mutagenesis in Nicotiana tabacum.
- Authors: Gao J, Wang G, Ma S, Xie X, Wu X, Zhang X, Wu Y, Zhao P, Xia Q
- Issue date: 2015 Jan
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