RNAi on progeny

Model Organism: Worms Methods & Reagents
1

Hi everybody,

I have a question regarding RNAi experiments. How do you do when you want to see an effect of RNAi on the progeny?
I usually do experiments on L1 worms and expose to RNAi for 2 or 3 days to see an effect on adults mainly. But I want to see an effect on young larvae so I’d like to treat worms to see the effect on the F1. I know that the maximum RNAi effect is usually observed after 48h exposure so I am wondering what the best way to do my experiments. I find it very convenient to use L1, but it might be too long if I treat L1 and wait until I can observe L1 or L2 of the next generation. Is it better to do the RNAi on L4?
It will be helpful for me if you could share your experience.
Thanks a lot!

2

Perhaps you can do RNAi by injection? This is much more labour intensive than RNAi by feeding though…

3

It’s feasible to perform RNAi feeding at any stage of development. Use age-synchronized cultures grown on OP50, then switch to RNAi plates at the desired time (wash to minimize OP50 transfer). It’s a good idea to try a time course to optimize the F1 phenotype.

-Harold

4

Thanks for your suggestions.
I’ll try first to do a time course by feeding and if it doesn’t give good results, injection seems a good idea.