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Polyelectrolyte Multilayer‐Mediated Gene Delivery for Semaphorin Signaling Pathway Control
Author(s) -
Richard Doriane,
Nguyen Isabelle,
Affolter Christine,
Meyer Florent,
Schaaf Pierre,
Voegel JeanClaude,
Bagnard Dominique,
Ogier Joelle
Publication year - 2010
Publication title -
small
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.785
H-Index - 236
eISSN - 1613-6829
pISSN - 1613-6810
DOI - 10.1002/smll.201000228
Subject(s) - semaphorin , polyelectrolyte , plasmid , gene delivery , transduction (biophysics) , microbiology and biotechnology , plexin , receptor , signal transduction , chemistry , biophysics , biology , dna , transfection , gene , biochemistry , organic chemistry , polymer
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.