Efficiency of gene transfection reagents in NG108-15, SH-SY5Y and CHO-K1 cell lines.
Several gene delivery reagents were analyzed for their transfection efficiency. Genes studied belonged to the class of mammalian proteins termed regulators of G-protein signaling (RGS), ranged in size up to 2.2 Kb long and were transfected into the NG108-15, SH-SY5Y and CHO-K1 cell lines. Prior to transfection, genes were cloned into a nonviral vector pcDNA 6.2/EmGFP, so as to express a green fluorescent protein tag at the 3' end. Flow cytometry was used to analyze cell fluorescent activity and thereby transfection efficiency. Gene delivery reagents Lipofectamine 2000 and ExGen 500 produced more effective transfection in NG108-15 cells whereas Lipofectamine 2000, ExGen 500 and TurboFectin 8.0 were more effective in CHO-K1 cells. In both these cell lines, transfection efficiency reached 60-80%. In SH-SY5Y cells, TurboFectin 8.0 produced the best transfection result; however efficiency level was only 5%. Gene size had no effect on transfection efficiency. Unlike Lipofectamine 2000, cells transfected using ExGen 500 showed morphological deformation. Our results suggest that Lipofectamine 2000 is the most suitable transfection medium for gene delivery to NG108-15 and CHO-K1 cells.
Martín-Montañez
E
E
Laboratory of Neurobiology, CIMES, Spain.
López-Téllez
J F
JF
Acevedo
M J
MJ
Pavía
J
J
Khan
Z U
ZU
D000818:Animals; D016466:CHO Cells; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D005910:Glioma / Q000235:genetics / Q000473:pathology; D006801:Humans; D008055:Lipids; D051379:Mice; D009447:Neuroblastoma / Q000235:genetics / Q000473:pathology; D051381:Rats; D014162:Transfection / Q000379:methods*
D008055:Lipids; C086724:Lipofectamine