Abstract
Polyethylenimine (PEI) has been used widely in transient gene expression studies of mammalian cells. We performed transient gene expression in suspension Chinese hamster ovary cells using a one-step transfection procedure in which DNA and PEI were simultaneously added to a cell culture in suspension without prior PEI/DNA complex incubation. To further understand the effect of PEI/DNA formation on the transfection and expression of exogenous gene in shaking state, we investigated the diameter and overcharge of the PEI/DNA complex. The results showed that the diameter of the complex was smaller with more positive charge when the PEI/DNA ratio was higher. Moreover, DNA more easily penetrated cells and nuclei at higher PEI concentrations. The highest transcription level, transfection efficiency, and GFP expression were obtained when the PEI/DNA ratio was 5:1.
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Acknowledgments
This study was supported by grants from the Guangdong Provincial project for cooperation of industry and academy (2010B090400500) and Major national science and technology project for New medicine research (2012ZX09202-301-001).
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Xie Qiuling and Guo Xinyong contributed equally to this work.
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Xie, Q., Xinyong, G., Xianjin, C. et al. PEI/DNA formation affects transient gene expression in suspension Chinese hamster ovary cells via a one-step transfection process. Cytotechnology 65, 263–271 (2013). https://doi.org/10.1007/s10616-012-9483-9
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DOI: https://doi.org/10.1007/s10616-012-9483-9