title: Polyelectrolyte multilayer-mediated gene delivery for semaphorin signaling pathway control.
abstract: The capability of multilayered polyelectrolyte films (MPFs) to control the sequential expression of two genes encoding cell receptors involved in a common cell signalling activity is shown, while achieving a fully functional signal transduction. As a functional model system representative of a cell signalling process that proceeds in a top-down manner, cell collapse induced by semaphorin 3A (Sema3A) was chosen as the target. Polyelectrolyte multilayers were sequentially functionalized with two plasmids encoding Neuropilin-1 (NRP-1) and Plexin-A1 (Plx-A1), respectively, acting as co-receptors for Sema3A. By using hyaluronan and chitosan as structural components for the incorporation of plasmid DNA layers onto precursor films made of poly-allylamine hydrochloride and poly-sodium-4-styrenesulfonate, the polyelectrolyte system is established; this systems is capable of delivering both plasmids to Cos-1 cells in a manner that permits control over the timing and the respective order in which the two plasmid DNA constructs are expressed. Importantly, it was observed that, following Sema3A stimulation, COS-1 cells co-expressing Plx-A1 and NRP-1 display a collapse phenotype, which is determined by the multilayer build-up scheme, and that the expression products of both transgenes embedded in MPFs are temporally functional over several days while acting their role of co-receptors for Sema3A.
authors.affiliation: INSERM UMR 977, 11, rue Humann, 67085 Strasbourg, France.
authors.forename: Doriane
authors.initials: D
authors.lastname: Richard
authors.forename: Isabelle
authors.initials: I
authors.lastname: Nguyen
authors.forename: Christine
authors.initials: C
authors.lastname: Affolter
authors.forename: Florent
authors.initials: F
authors.lastname: Meyer
authors.forename: Pierre
authors.initials: P
authors.lastname: Schaaf
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Voegel
authors.forename: Dominique
authors.initials: D
authors.lastname: Bagnard
authors.forename: Joelle
authors.initials: J
authors.lastname: Ogier
mesh_terms: D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002522:Chlorocebus aethiops; D018014:Gene Transfer Techniques; D007150:Immunohistochemistry; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D039942:Neuropilin-1 / Q000235:genetics / Q000378:metabolism; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry; D039961:Semaphorins / Q000494:pharmacology*; D015398:Signal Transduction / Q000235:genetics; D013451:Sulfonic Acids / Q000138:chemical synthesis / Q000737:chemistry; D014162:Transfection
chemical_list: D009419:Nerve Tissue Proteins; D011108:Polymers; D039961:Semaphorins; D013451:Sulfonic Acids; D039942:Neuropilin-1; C077114:styrenesulfonic acid polymer