siRNA transfection of non-adherent lymphoma cells - (Aug/31/2005 )
Has anyone had any luck with transfection of siRNA into non-adherent cell lines anything other than electroporation or viral vectors? I have tried using DMRIE-C (InVitrogen), siPORT amine (Ambion), siPORT NeoFX (Ambion), siPORT Lipid (Ambion), TransIT-TKO (Mirus) and have not been able to show knockdown using real-time quantitative PCR or Western blot. I've used fluorescently labeled GAPD siRNA (Ambion) and see a definite shift in fluorescence by flow cytometry with some of these agents but no reduction in expression with my assays. Can anyone help?
i have tried with classic methods like calcium-phosphate precipitation, PEI, liposome... and never obtained expected results
please, help us!
Before mechanical (electroporation, nucleofection) or viral systems (adeno, lenti), one reagent you should try is Gencarrier-2 for suspension cells (like Jurkat, primary lymphocytes). We have
some success using this reagent to knockdown STAT-5.