Abstract
Polyethylenimine has been used widely in transient gene expression with mammalian cells. To further understand its mediation of gene transfer, the transfection of HEK 293-F cells with dynamically prepared PEI/DNA complexes was studied with the help of fluorescent labeling. The efficiency of complex endocytosis/phagocytosis was found to correlate with the average sizes of complexes applied and complexes greater than 1 μm in diameter were likely excluded by the cells. Coupled with complex growth in size, the degree of association between PEI and DNA increased with the time of complex formation in the presence of competing ions. The blocking of transcription by complex formation necessitated complex dissociation in the nuclear environment for transcription to happen. Intracellularly, the fates of PEI complexed DNA therefore may be mostly determined by the degree of association. Results also suggested that the uptake of PEI/DNA complexes and subsequent protein expression were independent of the cell cycle stages of HEK 293-F cells.
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Acknowledgments
This project was funded by the National High Technology Program (National 863 Plan), Project No. 2007AA021702, the Ministry of Science & Technology, China. The authors wish to thank Mr. Zhao-feng Luo, the University of Science & Technology of China, Hefei, for his outstanding assistance in instrumental analyses.
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The authors Xiangzong Han and Qiangyi Fang contributed equally to this work.
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Han, X., Fang, Q., Yao, F. et al. The heterogeneous nature of polyethylenimine-DNA complex formation affects transient gene expression. Cytotechnology 60, 63–75 (2009). https://doi.org/10.1007/s10616-009-9215-y
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DOI: https://doi.org/10.1007/s10616-009-9215-y