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

Handle URI:
http://hdl.handle.net/10754/626046
Title:
Pea early-browning virus -mediated genome editing via the CRISPR/Cas9 system in Nicotiana benthamiana and Arabidopsis
Authors:
Ali, Zahir ( 0000-0002-7814-8908 ) ; Eid, Ayman; Ali, Shawkat ( 0000-0003-3282-3186 ) ; Mahfouz, Magdy M. ( 0000-0002-0616-6365 )
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.
KAUST Department:
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
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.
Publisher:
Elsevier BV
Journal:
Virus Research
Issue Date:
17-Oct-2017
DOI:
10.1016/j.virusres.2017.10.009
Type:
Article
ISSN:
0168-1702
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).
Additional Links:
http://www.sciencedirect.com/science/article/pii/S0168170217305543
Appears in Collections:
Articles

Full metadata record

DC FieldValue Language
dc.contributor.authorAli, Zahiren
dc.contributor.authorEid, Aymanen
dc.contributor.authorAli, Shawkaten
dc.contributor.authorMahfouz, Magdy M.en
dc.date.accessioned2017-10-30T08:39:52Z-
dc.date.available2017-10-30T08:39:52Z-
dc.date.issued2017-10-17en
dc.identifier.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.en
dc.identifier.issn0168-1702en
dc.identifier.doi10.1016/j.virusres.2017.10.009en
dc.identifier.urihttp://hdl.handle.net/10754/626046-
dc.description.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.en
dc.description.sponsorshipWe 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).en
dc.publisherElsevier BVen
dc.relation.urlhttp://www.sciencedirect.com/science/article/pii/S0168170217305543en
dc.subjectCRISPR/Cas9en
dc.subjectVirus-mediated Genome Engineeringen
dc.subjectVirus-mediated Genome Editingen
dc.subjectTargeted Mutagenesisen
dc.subjectTobacco rattle virusen
dc.subjectPea early browning virusen
dc.subjectArabidopsisen
dc.subjectNicotiana benthamianaen
dc.titlePea early-browning virus -mediated genome editing via the CRISPR/Cas9 system in Nicotiana benthamiana and Arabidopsisen
dc.typeArticleen
dc.contributor.departmentLaboratory for Genome Engineering, Division of Environmental and Biological Sciences and Engineering, 4700 King Abdullah University of Science and Technology, Thuwal, 23955-6900, Saudi Arabiaen
dc.identifier.journalVirus Researchen
kaust.authorAli, Zahiren
kaust.authorEid, Aymanen
kaust.authorAli, Shawkaten
kaust.authorMahfouz, Magdy M.en
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