Optimization of 25 kDa linear polyethylenimine for efficient gene delivery.
A 25-kDa linear polyethylenimine (25 kDa L-PEI) has proven to be efficient and versatile agent for gene delivery. Therefore, we determined the optimal transfection conditions of 25 kDa L-PEI and examined whether it has comparable transfection efficiency with other commercially available reagents, ExGen 500, LipofectAMINE 2000, and Effectene by using EGFP expression vector in different cell lines. Transfection efficiency and cytotoxicity were measured by flow cytometry. First of all, we determined the optimal ratio of nitrogen to phosphorous (N/P) and DNA concentration. With the increase of N/P ratio and DNA amounts, transfection efficiency increased with a slight variation in cell types. The optimal amounts of 25 kDa L-PEI were determined at N/P ratio 40 and DNA concentration varied among the cell types. In addition, 25 kDa L-PEI worked efficiently and was less toxic than other reagents. However, the efficiency and toxicity of all these reagents varied according to cell types as well as the ratio of DNA to reagents and the amounts of DNA. Our finding illustrates the importance of optimal transfection conditions of 25 kDa L-PEI to obtain maximal transgene expression with less cytotoxicity. Importantly, the optimization of those conditions may make possible to perform transfection cost-effectively and efficiently.
Huh
Sun-Hyung
SH
CHA Stem Cell Institute, Graduate School of Life Science and Biotechnology, Pochon CHA University, Kangnam-Gu, Seoul 135-907, South Korea.
Do
Hyun-Jin
HJ
Lim
Hye-Young
HY
Kim
Dong-Ku
DK
Choi
Seong-Jun
SJ
Song
Hyuk
H
Kim
Nam-Hyung
NH
Park
Jin-Ki
JK
Chang
Won-Kyong
WK
Chung
Hyung-Min
HM
Kim
Jae-Hwan
JH
D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D004274:DNA, Recombinant / Q000008:administration & dosage / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008970:Molecular Weight; D009584:Nitrogen / Q000032:analysis; D010758:Phosphorus / Q000032:analysis; D011094:Polyethyleneimine / Q000737:chemistry* / Q000633:toxicity; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection
D004274:DNA, Recombinant; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D010758:Phosphorus; D011094:Polyethyleneimine; D009584:Nitrogen