Abstract
Virus-induced gene silencing (VIGS) is an efficient tool for high throughput reverse genetic screens. VIGS engages the endogenous RNA-silencing machinery of the plant host, and can yield an 85–95% reduction of target transcripts. Gene silencing is rapid, target-specific, and does not require the creation of stable transformants. The technique has been used successfully in numerous Solanaceae species as well as in Arabidopsis, maize, and rice. Here we describe a protocol for conducting a VIGS screen in Nicotiana benthamiana using Tobacco Rattle Virus (TRV) based silencing vectors. This protocol can readily be adapted to many other model plant species.
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Notes
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Dimethylformamide is an eye and skin irritant, and is toxic to the liver and kidney. Wear appropriate safety equipment and dispense in a chemical fume hood.
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Acknowledgements
We thank past NSF funding in support of VIGS work in S, P. D-K lab.
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Hayward, A., Padmanabhan, M., Dinesh-Kumar, S.P. (2011). Virus-Induced Gene Silencing in Nicotiana benthamiana and Other Plant Species. In: Pereira, A. (eds) Plant Reverse Genetics. Methods in Molecular Biology, vol 678. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-682-5_5
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DOI: https://doi.org/10.1007/978-1-60761-682-5_5
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