chemical synthesized siRNA and rapidly dividing cells - (Dec/11/2004 )
I am new to RNAi area. Your suggestion and opinion are very important to me
If i am to use some rapidly dividing cells (e.g. HEK 293 cells) to perform RNAi study, is it not wise to use chemically synthesized siRNA?
I have this wonder because in this case the RNAi effects may be too insignificant to be detectable.
Is my concept correct? since i think for chemically synthesized siRNA, its RNAi signal cannot be replicate inside cells, and thus only cells transfected with siRNA will have effect. But new cells already grow if cells are rapidly dividing, leading to insignificant RNAi level.
Please let me know if my concept is correct. Thank you very much!
That may not always be the case. I have seen people knockdown genes in 293 and also in fast growing Hela cells using synthetic siRNAs.
For those fast growing cells, I usually do the transfection immediately after I plate the cells or a couple of hours after the cells have settled.
If you're worried about that you could use shRNA expression vectors, such as those we sell at http://www.expresson.com/productsandservices/vectors.html or many other people sell them too.