4 replies to this topic

[snc]

Hi
I am interested in using a vector based method to generate stable cell lines expressing RNAi targeted to a gene of interest.

Has anyone got any experience in using RNAi vectors to create stable cell lines?If so could you please recommend which vector I should use

Look forward to your reply
Thanks  

[rassen]

there is nothing special for stable expressing of siRNA as other genes. A lot of companies sell siRNA vectors with selection marker for stable expression such as Ambion and Invitrogen. I have bought the pSilencer CMV neo CMV-driven sirna expression vector from ambion but have not had time to insert my oligos into it. It is much cheaper than that sold by invitrogen but I don't know yet whether it works better or not.

[snc]

I bought the psilencer 4.1 cmv neo vector from Ambion as well. Chose it as most of the other vectors consist of the u6 promoter and this particular vector has been used quite a bit
Now its a matter of designing the shRNAi to clone in.
Thanks for the reply

[ahpaul]

the effect of siRNA is just a transient effect ... u can try 2 shift 2 shRNA which will express a stable cell line

[expresson_help]

Our vectors contain a neomycin marker for stable expression and are cheaper than a lot of competitors. More information at http://www.expresson...es/vectors.html

David
Expresson