Journalartikel

Jahr der Veröffentlichung: 2002

Seiten: 655-667

Zeitschrift: The Journal of Gene Medicine

Bandnummer: 4

Heftnummer: 6

ISSN: 1099-498X

eISSN: 1521-2254

DOI Link: https://doi.org/10.1002/jgm.313

Verlag: Wiley

Abstract: 

Background Several approaches for gene therapy of cystic fibrosis using viral and non-viral vectors are currently being undertaken. Nevertheless, the 3 present data suggest that vectors currently being used will either have to be further modified or, alternatively, novel vector systems need to be developed. 2 Recently, bacteria have been proven as suitable vehicles for DNA transfer to a wide variety of eukaryotic cells. In this study, we assessed the ability of the facultative intracellular pathogen Listeria monocytogenes to deliver a cDNA encoding the human cystic fibrosis transmembrane conductance regulator (CFTR) to CHO-K1 cells, since these cells have been extensively used for heterologous CFTR expression.

Methods An established in vitro gene transfer system based on antibiotic-mediated lysis of intracellular L. monocytogenes was exploited to transfer eukaryotic expression plasmids. Transient as well as stable CFTR transgene expression was analyzed by microscopical and biochemical methods; functionality was tested by whole-cell patch-clamp recordings.

Results L. monocytogenes mediated gene transfer to CHO-K1 cells was facilitated by an improved transfection protocol. In addition, the use of the isogenic mutant L. monocytogenes hlyW491A, engineered to produce a hemolysin variant with low toxigenic activity, greatly enhanced the efficiency of gene transfer. This strain allowed the transfer of functional CFTR to CHO-K1 cells.

Conclusions This is the first demonstration of L. monoyctogenes mediated CFTR transgene transfer. The successful in vitro transfer suggests that L. monocytogenes might be a potential vector for cystic fibrosis gene therapy or alternative applications and deserves further investigation in vitro as well as in vivo.

Harvard-Zitierstil: Krusch, S., Domann, E., Frings, M., Zelmer, A., Diener, M., Chakraborty, T., et al. (2002) Listeria monocytogenes mediated CFTR transgene transfer to mammalian cells, The Journal of Gene Medicine, 4(6), pp. 655-667. https://doi.org/10.1002/jgm.313

APA-Zitierstil: Krusch, S., Domann, E., Frings, M., Zelmer, A., Diener, M., Chakraborty, T., & Weiss, S. (2002). Listeria monocytogenes mediated CFTR transgene transfer to mammalian cells. The Journal of Gene Medicine. 4(6), 655-667. https://doi.org/10.1002/jgm.313