Guanidinium-grafted polyethylenimine: an efficient transfecting agent for mammalian cells.
Polyethylenimine (PEI) is one of the most efficient polycationic non-viral gene delivery vectors. Its efficiency and cytotoxicity depends on molecular weight, with the 25-kDa PEI being most efficient but accompanied with cytotoxicity. In the present study, enhancement in gene delivery efficiency along with reduction in cytotoxicity by attachment of guanidinium side group was explored. The hypothesis was that the guanidination would lead to the delocalization of charge present on primary amines of the polymer thereby leading to enhancement in gene delivery efficiency along with reduction in cytotoxicity. The polymer was guanidinated using O-methylisourea hemisulfate and the chemical linkage characterized by FTIR spectroscopy. The hydrodynamic diameter of guanidinated PEI-DNA complexes was determined using DLS. Subsequently, these complexes were used for DNA binding assay and zeta-potential measurements, taking native PEI as reference. Further, guanidinated PEI-DNA complexes were investigated for their gene delivery efficacy on HEK 293 cells. The hydrodynamic diameter of guanidinated PEI-DNA complexes was found to be in the range of 176-548 nm. As expected, the zeta potential values increased, on increasing the N/P ratios. It was found that guanidinated PEI had higher transfection efficiency at the majority of the N/P ratios tested as compared to commercially available transfecting agent lipofectin and native PEI itself. The toxicity of guanidinated PEI-DNA complexes was also reduced considerably in comparison to PEI polymer, as determined by MTT colorimetric assay. Out of the various derivatives prepared, gPEI 56% was found to be the most efficient in in vitro transfection.
Nimesh
Surendra
S
Dr. BR Ambedkar Center for Biomedical Research, University of Delhi, Delhi, India.
Chandra
Ramesh
R
D002478:Cells, Cultured; D004247:DNA / Q000008:administration & dosage*; D018014:Gene Transfer Techniques; D019791:Guanidine / Q000008:administration & dosage*; D006801:Humans; D010957:Plasmids; D011094:Polyethyleneimine / Q000008:administration & dosage*; D014162:Transfection / Q000379:methods*
D011094:Polyethyleneimine; D004247:DNA; D019791:Guanidine