Author/Editor		Tucker, Steven J; McClelland, David; Jaspars, Marcel; Sepčić, Kristina; MacEwan, David J; Scott, Roderick H
Title		The influence of alkyl pyridinium sponge toxins on membrane properties, cytotoxicity, transfection and protein expression in mammalian cells
Type		članek
Source		Biochim Biophys Acta
Vol. and No.		Letnik 1614
Publication year		2003
Volume		str. 171-81
Language		eng
Abstract		The ability of two alkyl pyridinium sponge toxin preparations (poly-APS and halitoxin) to form transient poresilesions in cell membranes and allow transfection of plasmid cDNA have been investigated using HEK 293 cells. Poly-APS and halitoxin preparations caused a collapse in membrane potential, reductions in input resistance and increased Caz + permeability. At least partial recovery was observed after poly-APS application but recovery was more rarely seen with halitoxin. The transfection with plasmid cDNAs for an enhanced green fluorescent protein (EGFP) and human tumour necrosis factor receptor 2 {TNFR2) was assessed for both toxin. preparations and compared with lipafectamine. Stable transfection was achieved with poly-APS although it was less efficient than lipofectamine. These results show that viable cells transfected with alien cDNA can be obtained using novel transient pore-forming a1ky1 pyridinium sponge toxins and a simple pre-incubation protocol. This provides the first proof of principle that pore-forming alkyl pyridinium compounds can be used to deliver cDNA to the intracellular environment without permanently compromising the plasma membrane.
Descriptors		PORIFERA TOXINS PYRIDINIUM COMPOUNDS CELL MEMBRANE PERMEABILITY CALCIUM TRANSFECTION TUMOR NECROSIS FACTOR DNA, COMPLEMENTARY PATCH-CLAMP TECHNIQUES FLUORESCENCE