First siRNA library screening in hard-to-transfect HUVEC cells.

摘要: Meaningful RNAi-based data for target gene identification are strongly dependent on the use of a biologically relevant cell type and efficient delivery highly functional siRNA reagents into selected type. Here we report Amaxa® Nucleofector® 96-well Shuttle® System screening in primary cells. Lonza’s Clonetics® HUVEC-Human Umbilical Vein Endothelial Cells were transfected with Thermo Scientific Dharmacon siGENOME® Libraries targeting protein kinases cycle related genes screened important viability. Of 37 hits, down-regulation 33 led to reduced proliferation or increased death, while two allowed better The validated four out 16 strongest hits (COPB2, PYCS, CDK4 MYC) influenced varying degrees, reflecting differing importance survival HUVEC Our results demonstrate that allows libraries types previously considered be difficult transfect. Thus, validation targets can now conducted cells, as selection is not limited those accessible by lipid-mediated transfection.