The influence of alkyl pyridinium sponge toxins on membrane properties, cytotoxicity, transfection and protein expression in mammalian cells

Steven John Tucker, David McClelland, Marcel Jaspars, K. Sepcic, David Joseph MacEwan, Roderick Hamilton Scott

Research output: Contribution to journalArticle

25 Citations (Scopus)

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 languageEnglish
Pages (from-to)171-181
Number of pages10
JournalBiochimica et Biophysica Acta (BBA) - Biomembranes
Volume1614
Issue number2
DOIs
Publication statusPublished - 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

Cite this

The influence of alkyl pyridinium sponge toxins on membrane properties, cytotoxicity, transfection and protein expression in mammalian cells. / Tucker, Steven John; McClelland, David; Jaspars, Marcel; Sepcic, K.; MacEwan, David Joseph; Scott, Roderick Hamilton.

In: Biochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 1614, No. 2, 2003, p. 171-181.

Research output: Contribution to journalArticle

Tucker, Steven John ; McClelland, David ; Jaspars, Marcel ; Sepcic, K. ; MacEwan, David Joseph ; Scott, Roderick Hamilton. / The influence of alkyl pyridinium sponge toxins on membrane properties, cytotoxicity, transfection and protein expression in mammalian cells. In: Biochimica et Biophysica Acta (BBA) - Biomembranes. 2003 ; Vol. 1614, No. 2. pp. 171-181.
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