Revisit complexation between DNA and polyethylenimine - Effect of uncomplexed chains free in the solution mixture on gene transfection
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摘要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 similar to 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. (C) 2010 Elsevier B.V. All rights reserved.
著者Yue YA, Jin F, Deng R, Cai JG, Chen YC, Lin MCM, Kung HF, Wu C
期刊名稱Journal of Controlled Release
詳細描述To ORKTS: DOI: 10.1016/j.jconrel.2010.10.028
出版年份2011
月份10
日期10
卷號155
期次1
出版社ELSEVIER SCIENCE BV
頁次67 - 76
國際標準期刊號0168-3659
電子國際標準期刊號1873-4995
語言英式英語
關鍵詞Cellular internalization; DNA condensation; Free polyethylenimine; Gene delivery; Laser light scattering
Web of Science 學科類別Chemistry; Chemistry, Multidisciplinary; CHEMISTRY, MULTIDISCIPLINARY; Pharmacology & Pharmacy; PHARMACOLOGY & PHARMACY

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