Mutating zebrafish, part 2

How can 'interfering RNA' be used to create fish models of human disease?

Before watching this video, see Part 1.

Transcript

Don Love (Auckland University): I wish it was simpler! It’s introducing into zebrafish’ embryos a short interfering RNA. So this thing we are introducing [a gene] would express something critical [the interfering RNA] which itself would lead to the degradation of the targeted transcript that is expressed by the zebrafish [the RNA of the affected gene which is associated with disease]. So we add something called A [the gene], which makes B [the interfering RNA]. B leads to the degradation of a particular gene transcript [the RNA of the affected gene which is associated with disease].

So the gene [associated with the disease] would still be transcribed, but with A leading to the synthesis of B. B would lead to degradation of that gene transcript. So the steady state levels of that transcript then plummet, and that’s your outcome. So my gene targeting is really transcript targeting, which you could measure. Because if the transcript is itself degraded, then we see no translation product - there is no protein

I’m adding A to make B, and I want A to make B under my control. So I want to add something to ensure that A is switched on. So the promoter driving A to express B has to be a regulated promoter. That is, I don’t want the promoter to be on all the time.