Transfection of SH-SY5Y cells - (Sep/17/2014 )

Hi All Anyone has experience in transient transfection in SH-SY5Y cells? I will need to perform transient transfection for downstream luciferase assay analysis. I have tried seeding the cells to 70% confluence following by standard lipofectamine2000 protocol. But there are only about 15% of cells expressing RFP 4 days after transfection. I'm welcoming any suggestions. Thank you!!!

Transient transfection using lipid-based techniques like LF2000 are unlikely to get you more than 30% transfection anyway, even in the easiest to transfect lines (e.g. 293, HeLa). If you need a higher transfection rate, consider using electroporation methods.

Thanks Bob! I have tried electroporation method with 10 ug DNA/ 100 mm petri dish, the RFP expression was very low (<10%) too.  

Thus far, beside the Lipo2000, I have also tried Genefactor and Novafector, all these transfection doesn't seems to work for SH-SY5Y cells even though the ratio and so had been change.

Anyone have experience transfecting SH-SY5Y using calcium phosphate method?

Some lines just don't transfect well.  That's all there is to it.  You might want to try the lipofectamine plus system or think about constructing an lentivirus.

Calcium phosphate systems tend to give poorer transfection than lipid based too.