Abstract
We have developed and optimized a method for delivering plasmid-based genes into the vertebrate brain. The method involves condensation of DNA into stable, small (<100 nm ø), and diffusible complexes using a cationic polymer, polyethylenimine (PEI). The approach is extremely versatile as it relies on use of plasmid, not viral-based constructions. This means that construction preparation and amplification is easy to carry out, risk free, and rapidly verified. The method is extremely efficient, giving very high yields for small (nanogram)quantities of plasmid. An overriding advantage is that it provides a convenient technique for studying physiological regulation of neuronal gene function within defined brain areas at defined developmental stages or in specific physiological states. One can thus obtain data on integrated transcriptional responses that, obviously, could not be obtained by an in vitro approach without resorting to germinal transgenesis. We have applied it to the analysis of protein function and promoter regulation in the central nervous systems (CNS) of mouse, rat, and Xenopus.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Abdallah, B., A. Hassan, C. Benoist, D. Goula, J.P. Behr, and B.A. Demeneix. 1996. A powerful nonviral vector for in vivo gene transfer into the adult mammalian brain: polyethylenimine. Hum. Gene Ther. 7:1947–1954.
Behr, J.P. and B.A Demeneix. 1998. Gene delivery with polycationic amphiphiles and polymers. Curr. Res. Mol. Ther. 1:5–12.
Boussif, O., F. Lezoualc’h, M.A. Zanta, M. Mergny, D. Scherman, B. Demeneix, and J.P. Behr. 1995. A novel, versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc. Nad. Acad. Sci. USA. 92:7297–7303.
Gérard, C., M.P. Martres, K. Lefévre, M.C. Miquel, D. Vergé, L. Lanfumey, E. Doucet, M. Hamon, and S. El Mestikawy. 1997. Immuno-localization of serotonin 5-HT6 receptor-like material in the rat central nervous system. Brain Res. 146:207–219.
Ghorbel, M.T., I. Seugnet, N. Hadj-Sahraoui, P. Topilko, G. Levi, and B.A. Demeneix. 1998. Thyroid hormone effects on Krox-24 transcription in the post-natal mouse brain are developmentally regulated but are not correlated with mitosis. Oncogene 18:917–924.
Goula, D., J. Remy, P. Erbacher, M. Wasowicz, G. Levi, B. Abdallah, and B. Demeneix. 1998a. Size, diffusibility and transfection performance of linear PEI/DNA complexes in the mouse central nervous system. Gene Ther. 5:712–717
Guissouma, H., M.T. Ghorbel, I. Seugnet, T. Ouatas, and B.A. Demeneix. 1998. Physiological regulation of hypothalamic TRH transcription in vivo is T3 receptor isoform-specific. FASEB J. 12:1755–1764.
Lehmann, A. 1974. Atlas Stéréotaxique du Cerveau de la Souris. CNRS Editions, Paris.
Martres, M.P., B.A. Demeneix, N. Hanoun, M. Hamon, and B. Giros. 1998. Up-and down-expression of dopamine transporter by plasmid DNA transfer in the rat brain. Eur. J. Neurosci. 10:3607–3616.
Sambrook, J., E.F Fritsch, and T. Maniatis. 1989. In N. Ford, C. Nolan, and M. Ferguson (Eds.), Molecular Cloning: A Laboratory Manual. CSH Laboratory Press, Cold Spring Harbor, NY.
Seed, B. and J.Y. Sheen. 1988. A simple phase extraction assay for chloramphenicol acetyl transferase. Gene 67:271–277.
Suh, J., H.-J. Paik, and B.K. Hwang. 1994. Ionization of polyethylenimine and polyallylamine at various pHs. Bioorg. Chem. 22:318–327.
Zhu, N., D. Liggitt, Y. Liu, and R. Debs. 1993. Systemic gene expression after intravenous DNA delivery into adult mice. Science 267:209–211.
Zlokarnik, G., P.A. Negulescu, T.E. Knapp, L. Mere, N. Burres, L. Feng, M. Whitney, K. Roemer, and R.Y. Tsien. 1998. Quantitation of transcription and clonal selection of single living cells with β-lactamase as reporter. Science 279:84–88.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2002 Springer-Verlag New York, Inc.
About this chapter
Cite this chapter
Demeneix, B.A., Lemkine, G.F., Guissouma, H. (2002). Polyethylenimine: A Versatile Cationic Polymer for Plasmid-Based Gene Delivery in the CNS. In: Merighi, A., Carmignoto, G. (eds) Cellular and Molecular Methods in Neuroscience Research. Springer, New York, NY. https://doi.org/10.1007/978-0-387-22460-2_4
Download citation
DOI: https://doi.org/10.1007/978-0-387-22460-2_4
Publisher Name: Springer, New York, NY
Print ISBN: 978-0-387-95386-1
Online ISBN: 978-0-387-22460-2
eBook Packages: Springer Book Archive