Disassembly of polyethylenimine-DNA particles in vitro: implications for polyethylenimine-mediated DNA delivery
Here a simple in vitro assay was used to investigate the disassembly of nanoparticles of polyethylenimine (PEI) and DNA. Particles were formed with various PEIs, allowed to mature for 10 min, and then exposed to different competitors (RNA, DNA, BSA or heparin) or to different conditions of pH or osmolarity. DNA release was determined by gel electrophoresis or spectroscopy. The presence of heparin or high salt yielded complete particle disassembly for all PEIs tested. The addition of RNA to particles formed with linear PEIs or branched 2 kDa PEI resulted in rapid DNA release, but RNA induced only partial disassembly of particles formed with large branched PEIs. In the presence of competitor DNA, slow disassembly was observed with particles made with linear PEIs or branched 2 kDa PEI but not for particles made with larger branched PEIs. The presence of BSA resulted in partial disassembly of PEI-DNA particles, but acidic pH did not affect particle stability. If particles were allowed to mature longer than 10 min in NaCl, subsequent heparin-mediated DNA release decreased as the incubation time and the PEI:DNA ratio increased. However, particles that matured in culture medium were disassembled by heparin independently of maturation time or PEI:DNA ratio. It was concluded that branched PEIs have a higher affinity for DNA than linear PEIs, that the intracellular disassembly of PEI-DNA particles may involve interactions between PEI and cellular RNA, and that extended maturation of PEI-DNA particles in NaCl prior to transfection may limit the intracellular release of plasmid DNA.
- URL: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17079047
Keywords: DNA/*administration & dosage/genetics ; Deoxyribonucleases/chemistry ; Drug Delivery Systems ; Electrophoresis ; Polyacrylamide Gel ; *Gene Transfer Techniques ; Heparin/chemistry ; Hydrogen-Ion Concentration ; Molecular Conformation ; Nanoparticles ; Osmolar Concentration ; Polyethyleneimine/*chemistry ; Proteins/chemistry/genetics ; RNA/chemistry ; Serum Albumin ; Bovine/chemistry
Laboratory of Cellular Biotechnology, Ecole Polytechnique Federale de Lausanne (EPFL), CH 1015 Lausanne, Switzerland.
Record created on 2007-06-05, modified on 2016-08-08