Hi All,
I’m helping a professor at my university update a genetics lab using C. elegans. One experiment we want to do involves RNAi repressors.
We want to start with gene Y, feed with RNAi for gene x, and end up with wild type. For instance, start with Unc, feed RNAi for Bli, then end up with WT.
Does anyone know if something like this is possible?
Thanks For The Help!
What about something like this: http://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.0030128&type=printable
Sean O’Rourke and Bruce Bowerman had a nice suppressor screen of dynein ts mutants. At the semi-permissive temp the ts mutants produced strong embryonic lethality, and they did an RNAi screen to look for genes whose inactivation restored embryonic viability.
Yes, it should definitely be possible and would commonly be called a ‘suppressor screen’ i.e you start with a phenotype and knock down additional genes to suppress that phenotype.
This may be of help: http://www.genetics.org/content/191/4/1031
A possible experiment is to use unc-54(r293), which is sensitive to nonsense mediated decay (NMD)
Strain: https://cgc.umn.edu/strain/CZ3086
(oddly, they don’t have the r293 single mutant. But you can get it from this strain.
RNAi probably isn’t as strong as the Smg mutant nulls (there are 7 genes). But maybe you can knock down smg-1 with RNAi such that it gives a qualitatively different phenotype.
There is also the smg-1 mutant, which completely suppresses the paralysis caused by r293
Strain: https://cgc.umn.edu/strain/TR1417
NMD is a pretty cool process for genome surveillance, so there’s that as a bonus.
A nice positive control for RNAi plates is pop-1(RNAi), which confers 100% lethality if the plates are good. We plate L4 animals, transfer after 24 hours, and the progeny from the transfer plate are 100% dead embryos.
Thank you all for the replies! These experiments are exactly what I was looking for.