Hi! I’m hoping anyone that has done undergrad RNAi screens might be able to help me with a list of your favorite RNAi targets that result in penetrant easily observable plate level RNAi phenotypes?
We use pop-1(RNAi) as our positive control. Everybody dies as embryo or L1.
unc-22 is supposed to work. Strong loss is the Twitcher phenotype, which is awesome. Weaker loss confers nicotine resistance.
Hi David,
In one of my lab classes I have used bli-1, dpy-11, and dpy-13 with good results.
Unc-22 has also worked. I use mek-2 as a positive control–embryonic lethality.
I recommend in addition to or instead of N2 you should use one or more Eri strains
from the stock center; they gave us nice results.
NL2099, rrf-3(pk1426)
YY216, eri-9(gg106)
GR1373, eri-1(mg366) (ts, Ste at 25C, recommend to maintain at 15C)
THANKS! We’re actually trying to test whether or not two overexpression lines we have generated are generally reducing RNAi sensitivity or doing something more interesting, so I wanted to compare directly to N2 rather than use rrf-3 or eri-1.
I have used dpy-10 and dpy-13 in my teaching lab. The nice thing about these is that you can quantify the changes by measuring the lengths. My (good) students are able to quantify the difference in sensitivity between N2 and eri-1
Also, you can get the bugs free from a Cold Spring Harbor site, http://www.silencinggenomes.org/
Janet