Virus-Induced Gene Silencing in Nicotiana benthamiana and Other Plant Species

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.