One of our featured articles for February comes from S.P. Dinesh-Kumar and colleagues at Yale University and is an update to a method first published a few years ago in our RNAi manual (now marked way down to a bargain price in both hardcover and paperback). RNAi has become a commonly-used tool for the down-regulation of genes in plants. The most effective means of accomplishing this gene silencing is through the use of viral vectors, with the Tobacco Rattle Virus (TRV) providing the most robust results. Virus-Induced Gene Silencing as a Tool for Delivery of dsRNA into Plants outlines a simple procedure for introducing TRV-based vectors into plants such as Arabidopsis, Nicotiana benthamiana and tomato.
Like all of our featured articles, access to this protocol is free for both subscribers and non-subscribers alike.
June 20, 2010 at 3:58 pm
Having talked about the utility of VIGS with many individuals using it, the feeling seems to be that is an extremely ineffcient way of triggering RNAi in plants, mostly because of the mosaic nature of dsRNA expression (which follows viral spread). I think the comments above are slightly misleading…let’s
not forget, one of our best guesses for the origin of ‘classical’ RNAi is as an anti-viral response. Loading plant cells with viral RNA just leads
to expression peaks in RNAi inhibitors, and a reduction in efficacy.
June 21, 2010 at 8:20 am
Thanks for the comment Jonny. I think you’re right, in that what I’ve written above is a bit ambiguous. The use of TRV is the most efficient of VIGS vectors, not necessarily the most efficient means of gene silencing altogether, sorry if I was unclear. Please do take a look at the actual protocol (it’s one of our featured articles and is freely accessible to subscribers and non-subscribers alike). The authors make a more eloquent case for VIGS than I can, given their higher level of expertise. Each protocol has space for comments (see Update/Discuss This Protocol in the info box at top right). If you have suggestions for readers of alternative techniques, I’m sure they would be greatly appreciated (and of course we’d welcome submission of alternate protocols as well).