ABSTRACT
An acid labile branched PDMAEMA/acetal copolymer with amino group was synthesized by the DE-ATRP and followed by Michael addition. The degradation of the polymer was strongly pH-dependent. High nucleic acid transfection efficiency with low cytotoxicity was observed compared to its non-degradable copolymer counterpart.
Subject(s)
Acetals/chemistry , DNA/administration & dosage , Methacrylates/chemistry , Nylons/chemistry , Plasmids/administration & dosage , Transfection , Acetals/chemical synthesis , Adipose Tissue/cytology , Animals , Cell Line , DNA/genetics , Hydrolysis , Luciferases/genetics , Methacrylates/chemical synthesis , Nylons/chemical synthesis , Plasmids/genetics , Rabbits , Stem Cells/metabolismABSTRACT
The purpose of this study was to develop a platform transfection technology, for applications in the brain, which could transfect astrocytes without requiring cell specific functionalization and without the common cause of toxicity through high charge density. Here we show that a simple and scalable preparation technique can be used to produce a "knot" structured cationic polymer, where single growing chains can crosslink together via disulphide intramolecular crosslinks (internal cyclizations). This well-defined knot structure can thus "untie" under reducing conditions, showing a more favorable transfection profile for astrocytes compared to 25 kDa-PEI (48-fold), SuperFect® (39-fold) and Lipofectamine®2000 (18-fold) whilst maintaining neural cell viability at over 80% after four days of culture. The high transfection/lack of toxicity of this knot structured polymer in vitro, combined with its ability to mediate luciferase transgene expression in the adult rat brain, demonstrates its use as a platform transfection technology which should be investigated further for neurodegenerative disease therapies.
