CRISPR-Cas12a Genome Editing at the Whole-Plant Level Using Two Compatible RNA Virus Vectors
| Main Authors: | Uranga, Mireia, Vazquez-Vilar, Marta, Orzáez, Diego, Darós, José-Antonio |
|---|---|
| Format: | Article Journal |
| Bahasa: | eng |
| Terbitan: |
, 2021
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| Online Access: |
https://zenodo.org/record/5932122 |
Daftar Isi:
- The use of viral vectors that can replicate and move systemically through the host plant to deliver bacterialCRISPR components enables genome editing at the whole-plant level and avoids the requirement for labor-intensive stable transformation. However, this approach usually relies on previously transformed plants thatstably express a CRISPR-Cas nuclease. Here, we describe successful DNA-free genome editing ofNicotiana ben-thamianausing two compatible RNA virus vectors derived from tobacco etch virus (TEV; genusPotyvirus) andpotato virus X (PVX; genusPotexvirus), which replicate in the same cells. The TEV and PVX vectors respectivelyexpress a Cas12a nuclease and the corresponding guide RNA. This novel two-virus vector system improvesthe toolbox for transformation-free virus-induced genome editing in plants and will advance efforts to breedmore nutritious, resistant, and productive crops.
- This research was supported by grants BIO2017-83184-Rand PID2019-108203RB-I00 from the Ministerio deCiencia e Innovacio ́n (Spain) through the Agencia Estatalde Investigacio ́n (co-financed by the European RegionalDevelopment Fund) and H2020-760331 Newcotiana fromthe European Commission. M.U. and M.V.-V. are the recip-ients of fellowships FPU17/05503 from the Ministerio deCiencia e Innovacio ́n (Spain) and APOSTD/2020/096from the Generalitat Valenciana (Spain), respectively.