Repositorium
Pea early-browning virus -mediated genome editing via the CRISPR/Cas9 system in Nicotiana benthamiana and Arabidopsis
276
Journal Article / 2018
Ali, Zahir; Eid, Ayman; Ali, Shakila; Mahfouz, Magdy M.
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.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
666 |
Mahfouz, Magdy M. Saudi Arabia |
Arabidopsis thaliana |
CRISPR/Cas9 GL1 |
Lack trichomes (easily visible phenotypes) SDN1 |
Basic research Basic research |
667 |
Mahfouz, Magdy M. Saudi Arabia |
Arabidopsis thaliana |
CRISPR/Cas9 TT4 |
Transparent testa (easily visible phenotypes) SDN1 |
Basic research Basic research |
668 |
Mahfouz, Magdy M. Saudi Arabia |
Nicotiana benthamiana |
CRISPR/Cas9 PDS |
Albino phenotype SDN1 |
Basic research Basic research |