Abstract
The ability of two alkyl pyridinium sponge toxin preparations (poly-APS and halitoxin) to form transient pores/lesions 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 Ca2+ 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 turnout necrosis factor receptor 2 (TNFR2) was assessed for both toxin preparations and compared with lipofectamine. 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 alkyl 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. (C) 2003 Elsevier B.V. All rights reserved.
Original language | English |
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Pages (from-to) | 171-181 |
Number of pages | 10 |
Journal | Biochimica et Biophysica Acta (BBA) - Biomembranes |
Volume | 1614 |
Issue number | 2 |
DOIs | |
Publication status | Published - 2003 |
Keywords
- transfection
- pore formation
- sponge toxin
- cDNA
- protein expression
- GREEN-FLUORESCENT PROTEIN
- FACTOR RECEPTOR SUBTYPES
- TUMOR-NECROSIS
- 3-ALKYLPYRIDINIUM POLYMERS
- RENIERA-SARAI
- PORE
- CHANNEL
- DNA
- HALITOXIN
- COMPLEXES