Multilayered polyelectrolyte films promote the direct and localized delivery of DNA to cells.
Multilayered polyelectrolyte films fabricated from plasmid DNA and a hydrolytically degradable synthetic polycation can be used to direct the localized transfection of cells without the aid of a secondary transfection agent. Multilayered assemblies 100 nm thick consisting of alternating layers of synthetic polymer and plasmid DNA encoding for enhanced green fluorescent protein (EGFP) were deposited on quartz substrates using a layer-by-layer fabrication procedure. The placement of film-coated slides in contact with COS-7 cells growing in serum-containing culture medium resulted in gene expression in cells localized under the film-coated portion of the slides. The average percentage of cells expressing EGFP relative to the total number of cells ranged from 4.6% to 37.9%, with an average of 18.6%+/-8.2%, as determined by fluorescence microscopy. In addition to providing a mechanism for the immobilization of DNA at the cell/surface interface, a preliminary analysis of film topography by atomic force microscopy (AFM) demonstrated that polymer /DNA films undergo significant structural rearrangements upon incubation to present surface bound condensed plasmid DNA nanoparticles. These data suggest that the presence of the cationic polymer in these materials may also contribute to the internalization and expression of plasmid. The materials and design principles reported here present an attractive framework for the local or non-invasive delivery of DNA from the surfaces of implantable materials or biomedical devices.
Department of Chemical and Biological Engineering, University of Wisconsin - Madison, 1415 Engineering Drive, Madison, Wisconsin 53706-1607, USA.
Christopher M
CM
Jewell
Jingtao
J
Zhang
Nathaniel J
NJ
Fredin
David M
DM
Lynn
D000818:Animals; D019556:COS Cells; D002626:Chemistry, Pharmaceutical; D002522:Chlorocebus aethiops; D004337:Drug Carriers; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000008:administration & dosage* / Q000737:chemistry; D011094:Polyethyleneimine / Q000737:chemistry; D011108:Polymers / Q000008:administration & dosage* / Q000138:chemical synthesis* / Q000737:chemistry; D011791:Quartz; D013451:Sulfonic Acids / Q000737:chemistry; D014162:Transfection / Q000379:methods*; D000097573:Red Fluorescent Protein
D004337:Drug Carriers; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011094:Polyethyleneimine; D011108:Polymers; D011791:Quartz; D013451:Sulfonic Acids; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein; C077114:styrenesulfonic acid polymer