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 w...

Full description

Saved in:
Bibliographic Details
Published in:Journal of controlled release 2011-10, Vol.155 (1), p.67-76
Main Authors: Yue, Yanan, Jin, Fan, Deng, Rui, Cai, Jinge, Chen, Yangchao, Lin, Marie C.M., Kung, Hsiang-Fu, Wu, Chi
Format: Article
Language:English
Subjects:
Cell Line
Cellular internalization
Condensation
Controlled release
DNA
DNA - administration & dosage
DNA - metabolism
DNA condensation
Flow Cytometry
Free polyethylenimine
Gel electrophoresis
Gene delivery
genes
Humans
Kinetics
Laser light scattering
Lasers
Light scattering
Models, Molecular
Nitrogen
Phosphate
Polyethyleneimine - metabolism
Scattering, Radiation
Transfection
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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. [Display omitted]
ISSN:0168-3659
1873-4995
DOI:10.1016/j.jconrel.2010.10.028