Revisit complexation between DNA and polyethylenimine - Effect of uncomplexed chains free in the solution mixture on gene transfection.
Our revisit of the complexation between anionic DNA and cationic polyethylenimine (PEI) in both water and phosphate buffered saline (PBS) by using a combination of laser light scattering (LLS) and gel electrophoresis confirms that nearly all the DNA chains are complexed with PEI to form polyplexes when the molar ratio of nitrogen from PEI to phosphate from DNA (N:P) reaches ~3, but the PEI/DNA polyplexes have a high in-vitro gene transfection efficiency only when N:P≥10. Putting these two facts together, we not only conclude that this extra 7 portions of PEI chains are free in the solution mixture, but also confirmed that it is these free PEI chains that substantially promote the gene transfection no matter whether they are applied hours before or after the administration of the much less effective PEI/DNA polyplexes (N:P=3). The uptake kinetics measured by flow cytometry shows that the addition of free PEI leads to a faster and more efficient cellular internalization of polyplexes, but these free PEI chains mainly contribute to the subsequent intracellular trafficking. In contrast, the bound PEI chains mainly play a role in the DNA condensation and protection, leading to a different thinking in the development of non-viral vectors.
Department of Chemistry, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong. yananyue.linda@gmail.com
Yanan
Y
Yue
Fan
F
Jin
Rui
R
Deng
Jinge
J
Cai
Yangchao
Y
Chen
Marie C M
MC
Lin
Hsiang-Fu
HF
Kung
Chi
C
Wu
D002460:Cell Line; D004247:DNA / Q000008:administration & dosage* / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D007834:Lasers; D008958:Models, Molecular; D011094:Polyethyleneimine / Q000378:metabolism*; D012542:Scattering, Radiation; D014162:Transfection
D011094:Polyethyleneimine; D004247:DNA