Polynorbornene polycationic polymers as gene transfer agents. Influence of the counterion for in vitro transfection.
Polycationic derivatives of polynorbornene with different non-cytotoxic counterions, have been prepared by organometallic polymerization of methyleneammonium norbornene and subsequent exchange of the counterion. In this paper the effect of the counterion on the polycationic polymer binding onto plasmid DNA was studied via different ethidium bromide assays, heparin displacement and protection against degradation by DNAse. According to the nature of the counterions and consequently the size of the polymer particles, their complexation with the DNA led to aggregates with variable binding affinity for the plasmid. The relative transfection efficiency of each polyplex was compared, on the basis of reporter gene expression, in cells in culture. The nature of the counterion was seen to affect gene delivery. The order of transfection efficiency of the counterions studied at equivalent charge ratios (NH3+/PO4-) is lactobionate, acetate, chloride. The results obtained with the polynorbornene methyleneammonium lactobionate and acetate are particularly encouraging.
Laboratoire des I.M.R.C.P. UMR 5623 CNRS, Université Paul Sabatier, 118 route de Narbonne, 31062 Toulouse Cedex 04, France.
Saâdia
S
Asgatay
Sophie
S
Franceschi-Messant
Emile
E
Perez
Patricia
P
Vicendo
Isabelle
I
Rico-Lattes
Emilie
E
Phez
Marie Pierre
MP
Rols
D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D000818:Animals; D016466:CHO Cells; D002412:Cations / Q000737:chemistry / Q000494:pharmacology; D006224:Cricetinae; D003412:Cricetulus; D004249:DNA Damage / Q000187:drug effects / Q000502:physiology; D003850:Deoxyribonuclease I / Q000378:metabolism; D004187:Disaccharides / Q000737:chemistry / Q000145:classification; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004996:Ethidium / Q000378:metabolism; D017930:Genes, Reporter / Q000187:drug effects / Q000502:physiology; D005822:Genetic Vectors / Q000138:chemical synthesis / Q000494:pharmacology*; D006493:Heparin / Q000378:metabolism; D036103:Nanotechnology / Q000379:methods; D010316:Particle Size; D010957:Plasmids / Q000235:genetics; D010969:Plastics / Q000138:chemical synthesis / Q000145:classification* / Q000378:metabolism*; D011108:Polymers / Q000138:chemical synthesis / Q000494:pharmacology; D013678:Technology, Pharmaceutical / Q000379:methods; D014162:Transfection / Q000379:methods*
D002412:Cations; D004187:Disaccharides; D010969:Plastics; D011108:Polymers; C071464:polynorbornen; C005608:lactobionic acid; D006493:Heparin; D003850:Deoxyribonuclease I; D004996:Ethidium