Evaluation of transfection methods for transient gene expression in Chinese hamster ovary cells

DOI: 10.4236/abb.2013.412135   PDF   HTML     6,587 Downloads   9,651 Views   Citations

Abstract

Three transfection reagents, Lipofectamine® 2000, TransIT-PRO® and linear 25 kDa polyethylenimine were evaluated for transient expression of enhanced green fluorescent protein in Chinese hamster ovary cells. TransIT-PRO® was found to be more efficient under the examined conditions, but comes at an increased cost compared to the widely used PEI.

Share and Cite:

Sou, S. , Polizzi, K. and Kontoravdi, C. (2013) Evaluation of transfection methods for transient gene expression in Chinese hamster ovary cells. Advances in Bioscience and Biotechnology, 4, 1013-1019. doi: 10.4236/abb.2013.412135.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] Codamo, J., Munro, T.P., Hughes, B.S., Song, M. and Gray, P.P. (2011) Enhanced CHO cell-based transient gene expression with the Epi-CHO expression system. Molecular Biotechnology, 48, 109-115.
[2] Morrow, K.J. (2008) Optimizing transient gene expression. Genetic Engineering & Biotechnology News, 28, 54-59.
[3] Ehrhardt, C., Schmolke, M., Matzke, A., Knoblauch, A., Will, C., Wixler, V. and Ludwig S. (2006) Polyethylenimine, a cost-effective transfection reagent. Signal Transduction, 6, 179-184.
http://dx.doi.org/10.1002/sita.200500073
[4] Zaruc, V., Weltin, D., Erbacher, P., Remy, J.S., Behr, J.P. and Stephan, D. (2004) Effective polyethlenimine-mediated gene transfer into hunman endothelial cells. The Journal of Gene Medicine, 6, 176-184.
http://dx.doi.org/10.1002/jgm.484
[5] Obata, Y., Ciofani, G., Raffa, V., Cuschieri, A., Menciassi, A., Dario, P. and Takeoka, S. (2010) Evaluation of cationic liposomes composed of an amino acid-based lipid for neuronal transfection. Nanomedicine: Nanotechnology, Biology and Medicine, 6, 70-77.
http://dx.doi.org/10.1016/j.nano.2009.04.005
[6] Juckem, K.L. (2010) Maximize protein expression in CHO suspension cells. BioProcess International, 8, 122-123.
[7] Galbraith, D.J., Tait, A.S., Racher, A.J., Birch, J.R. and James, D.C. (2006) Control of culture environment for improved polyethylenimine-mediated transient production of recombinant monoclonal antibodies by CHO cells. Biotechnology Progress, 22, 753-762.
http://dx.doi.org/10.1021/bp050339v
[8] Thompson, B.C., Segarra, C.R.J., Mozley, O.L., Daramola, O., Field, R., Levison, P.R. and James, D.C. (2012) Cell line specific control of polyethyleniminemediated transient transfection optimized with “Design of experiments” methodology. Biotechnology Progress, 28, 179-187. http://dx.doi.org/10.1002/btpr.715
[9] Xie, Q.L., Guo, X.Y., Chen, X.J. and Wang, Y.Y. (2013) PEI/DNA formation affects transient gene expression in suspension Chinese hamster ovary cells via a one-step transfection process. Cytotechnology, 65, 263-271.
http://dx.doi.org/10.1007/s10616-012-9483-9
[10] Wulhfard, S., Tissot, S., Bouchet, S., Cevey, J., De Jesus, M., Hacker, D.L. and Wurm, F.M. (2008) Mild hypothermia improves transient gene expression yields several fold in chinese hamster ovary cells. Biotechnology Progress, 24, 458-465.
http://dx.doi.org/10.1021/bp070286c
[11] Roobol, A., Carden, M.J., Newsam, R.J. and Smales, C.M. (2009) Biochemical insights into the mechanisms central to the response of mammalian cells to cold stress and subsequent rewarming. FEBS Journal, 276, 286-302.
http://dx.doi.org/10.1111/j.1742-4658.2008.06781.x
[12] Masterton, R.J., Roobol, A., Al-Fageeh, M.B., Carden, M.J. and Smales, C.M. (2010) Post translation events of a model reporter protein proceed with higher fidelity and accuracy upon mild hypothermic culturing of Chinese Hamster Ovary cells. Biotechnology and Bioengineering, 105, 215-220.
http://dx.doi.org/10.1002/bit.22533

  
comments powered by Disqus

Copyright © 2020 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.