Transfection of B-cells - siRNA transfection in Primary effusion lymphomas (Feb/01/2008 )

Hi,
Anyone have experience to transfect B cells. I want to use siRNA transfection in Primary effusion lymphoma e.g. BCBL1, BC1, BC3 cells. I have Ambion pSilencer adenoviral expression system but it doesn’t have infectability to transfect B-cells. Please suggest me an efficient system to transfect these cells.

Thanks

You may want to consider a retroviral vector. A girl in the lab next door has used the clontech pSiren RetroQ Green vector to knock down expression of her gene of interest in B cells:

http://www.clontech.com/images/pt/PT3777-5.pdf

We also use retroviral vectors for all the genes we wish to over-express in primary hematopoietic cells.

Thank you "stemcell69" for your suggestion. I think none of the system can give efficient delivery of siRNA into B cells (non adherence cells) as compared to other adherence cell lines. However, some researchers successfully transduce siRNA into Human B cells by retrovirus vectors/ lentivirus. I am planning to use retrovirus vector.

Anybody can suggest me which retrovirus vector is good for siRNA delivery into B cell, either pSUPER.retro.puro (Oligoengine) OR pSIREN-Retro Q-ZsGreenwhich vector (clontech) and what the importance to choose promoter U6 and H1.

Thanks

well the difference of price between the 2 vectors seems to be in favor of buying pSIREN RETRO Q GREEN vector.
In fact you may be cell sorting the cells expressing the GFP which is a big advantage with hard-to-transfect cells.

U6 or H1 are similar in terms of activity for expressing shRNAs in human cells.

I don't know if you still need to know this, but I use HiPerFect to transfect siRNA into myeloma cell lines. I get ~95% positive cells at 24h.