Influence of macromolecular architecture on nucleic acid transfection

POLY 611

John M. Layman1, Anjali A. Hirani2, Joseph M. Pickel3, Phillip F. Britt, brittpf@ornl.gov4, Yong Woo Lee, ywlee@vt.edu2, and Timothy E. Long, telong@vt.edu5. (1) Department of Chemistry, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, (2) School of Biomedical Engineering and Sciences, Virginia Tech, 1121 Research Building XV, 1880 Pratt Drive, Blacksburg, VA 24061, (3) Chemical Sciences Division, Oak Ridge National Laboratory, One Bethel Valley Road, P.O.Box 20, Oak Ridge, TN 37831, (4) Center for Nanophase Materials Sciences and Chemical Science Division, Oak Ridge National Laboratory, Building 4500S, S-144, MS-6129, P.O. Box 2008, Oak Ridge, TN 37831-6197, (5) Department of Chemistry, Virginia Tech, 124 Davidson Hall, Blacksburg, VA 24061
Non-viral gene transfection agents, such as cationic polyelectrolytes, are attractive replacements to viruses due to the absence of potential immunogenic risk and the ability to tailor their macromolecular architecture. In this study, conventional free radical polymerization was used to synthesize various molecular weights of linear and randomly branched cationic polyelectrolytes based on poly(2-N,N'-dimethylaminoethyl methacrylate)•hydrochloride (PDMAEMA•HCl). Randomly branched topologies were synthesized utilizing an ethylene glycol based dimethacrylate as a branching agent. Our work has focused on the influence of the degree of branching on transfection efficiency and toxicity in human brain microvascular endothelial (HBMEC) cells. We described the degree of branching using the well established, semi-quantitative, g' value, which is the ratio of intrinsic viscosities of branched and linear polymers at similar molar mass. Cell viability assays found limited cytotoxicity of various molecular weights of linear and branched PDMAEMA•HCl complexed with the pRL-SV40 Renilla luciferase expression plasmid over the concentrations, times, and cell type tested. Futhermore, the topology of the PDMAEMA•HCl gene transfer agent was found to have a significant effect on gene transfection in HBMEC cells.
 

7th International Biorelated Polymers Symposium
1:30 PM-5:20 PM, Wednesday, 13 September 2006 San Francisco Marriott -- Salon 14/15, Oral

Division of Polymer Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006