Pore forming polyalkylpyridinium salts from marine sponges versus synthetic lipofection systems

distinct tools for intracellular delivery of cDNA and siRNA

Debra McLaggan, Noppadon Adjimatera, Kristina Sepcic, Marcel Jaspars, David J MacEwan, Ian S Blagbrough, Roderick H Scott

Research output: Contribution to journalArticle

17 Citations (Scopus)
4 Downloads (Pure)

Abstract

Background: Haplosclerid marine sponges produce pore forming polyalkylpyridinium salts ( poly-APS), which can be used to deliver macromolecules into cells. The aim of this study was to investigate the delivery of DNA, siRNA and lucifer yellow into cells mediated by poly-APS and its potential mechanisms as compared with other lipofection systems ( lipofectamine and N-4, N(9-)dioleoylspermine (LipoGen)). DNA condensation was evaluated and HEK 293 and HtTA HeLa cells were used to investigate pore formation and intracellular delivery of cDNA, siRNA and lucifer yellow.

Results: Poly-APS and LipoGen were both found to be highly efficient DNA condensing agents. Fura-2 calcium imaging was used to measure calcium transients indicative of cell membrane pore forming activity. Calcium transients were evoked by poly-APS but not LipoGen and lipofectamine. The increases in intracellular calcium produced by poly-APS showed temperature sensitivity with greater responses being observed at 12 C compared to 21 C. Similarly, delivery of lucifer yellow into cells with poly-APS was enhanced at lower temperatures. Transfection with cDNA encoding for the expression enhanced green fluorescent protein was also evaluated at 12 C with poly-APS, lipofectamine and LipoGen. Intracellular delivery of siRNA was achieved with knockdown in beta-actin expression when lipofectamine and LipoGen were used as transfection reagents. However, intracellular delivery of siRNA was not achieved with poly-APS.

Conclusion: Poly-APS mediated pore formation is critical to its activity as a transfection reagent, but lipofection systems utilise distinct mechanisms to enable delivery of DNA and siRNA into cells.

Original languageEnglish
Article number6
Number of pages12
JournalBMC Biotechnology
Volume6
DOIs
Publication statusPublished - 16 Jan 2006

Fingerprint

Porifera
Small Interfering RNA
Complementary DNA
Salts
Calcium
Transfection
DNA
Temperature
Fura-2
HeLa Cells
Actins
Cell Membrane
Lipofectamine
lucifer yellow

Keywords

  • gene delivery
  • Reniera-sarai
  • membrane-properties
  • Haliclona-vicosa
  • ethidium-bromide
  • mammalian-cells
  • DNA
  • spermine
  • condensation
  • transfection

Cite this

Pore forming polyalkylpyridinium salts from marine sponges versus synthetic lipofection systems : distinct tools for intracellular delivery of cDNA and siRNA. / McLaggan, Debra; Adjimatera, Noppadon; Sepcic, Kristina; Jaspars, Marcel; MacEwan, David J; Blagbrough, Ian S; Scott, Roderick H.

In: BMC Biotechnology, Vol. 6, 6, 16.01.2006.

Research output: Contribution to journalArticle

McLaggan, Debra ; Adjimatera, Noppadon ; Sepcic, Kristina ; Jaspars, Marcel ; MacEwan, David J ; Blagbrough, Ian S ; Scott, Roderick H. / Pore forming polyalkylpyridinium salts from marine sponges versus synthetic lipofection systems : distinct tools for intracellular delivery of cDNA and siRNA. In: BMC Biotechnology. 2006 ; Vol. 6.
@article{5e245b1f804f4f20acf1c1e1f5f8e33e,
title = "Pore forming polyalkylpyridinium salts from marine sponges versus synthetic lipofection systems: distinct tools for intracellular delivery of cDNA and siRNA",
abstract = "Background: Haplosclerid marine sponges produce pore forming polyalkylpyridinium salts ( poly-APS), which can be used to deliver macromolecules into cells. The aim of this study was to investigate the delivery of DNA, siRNA and lucifer yellow into cells mediated by poly-APS and its potential mechanisms as compared with other lipofection systems ( lipofectamine and N-4, N(9-)dioleoylspermine (LipoGen)). DNA condensation was evaluated and HEK 293 and HtTA HeLa cells were used to investigate pore formation and intracellular delivery of cDNA, siRNA and lucifer yellow. Results: Poly-APS and LipoGen were both found to be highly efficient DNA condensing agents. Fura-2 calcium imaging was used to measure calcium transients indicative of cell membrane pore forming activity. Calcium transients were evoked by poly-APS but not LipoGen and lipofectamine. The increases in intracellular calcium produced by poly-APS showed temperature sensitivity with greater responses being observed at 12 C compared to 21 C. Similarly, delivery of lucifer yellow into cells with poly-APS was enhanced at lower temperatures. Transfection with cDNA encoding for the expression enhanced green fluorescent protein was also evaluated at 12 C with poly-APS, lipofectamine and LipoGen. Intracellular delivery of siRNA was achieved with knockdown in beta-actin expression when lipofectamine and LipoGen were used as transfection reagents. However, intracellular delivery of siRNA was not achieved with poly-APS. Conclusion: Poly-APS mediated pore formation is critical to its activity as a transfection reagent, but lipofection systems utilise distinct mechanisms to enable delivery of DNA and siRNA into cells.",
keywords = "gene delivery, Reniera-sarai, membrane-properties, Haliclona-vicosa, ethidium-bromide, mammalian-cells, DNA, spermine, condensation, transfection",
author = "Debra McLaggan and Noppadon Adjimatera and Kristina Sepcic and Marcel Jaspars and MacEwan, {David J} and Blagbrough, {Ian S} and Scott, {Roderick H}",
year = "2006",
month = "1",
day = "16",
doi = "10.1186/1472-6750-6-6",
language = "English",
volume = "6",
journal = "BMC Biotechnology",
issn = "1472-6750",
publisher = "BioMed Central",

}

TY - JOUR

T1 - Pore forming polyalkylpyridinium salts from marine sponges versus synthetic lipofection systems

T2 - distinct tools for intracellular delivery of cDNA and siRNA

AU - McLaggan, Debra

AU - Adjimatera, Noppadon

AU - Sepcic, Kristina

AU - Jaspars, Marcel

AU - MacEwan, David J

AU - Blagbrough, Ian S

AU - Scott, Roderick H

PY - 2006/1/16

Y1 - 2006/1/16

N2 - Background: Haplosclerid marine sponges produce pore forming polyalkylpyridinium salts ( poly-APS), which can be used to deliver macromolecules into cells. The aim of this study was to investigate the delivery of DNA, siRNA and lucifer yellow into cells mediated by poly-APS and its potential mechanisms as compared with other lipofection systems ( lipofectamine and N-4, N(9-)dioleoylspermine (LipoGen)). DNA condensation was evaluated and HEK 293 and HtTA HeLa cells were used to investigate pore formation and intracellular delivery of cDNA, siRNA and lucifer yellow. Results: Poly-APS and LipoGen were both found to be highly efficient DNA condensing agents. Fura-2 calcium imaging was used to measure calcium transients indicative of cell membrane pore forming activity. Calcium transients were evoked by poly-APS but not LipoGen and lipofectamine. The increases in intracellular calcium produced by poly-APS showed temperature sensitivity with greater responses being observed at 12 C compared to 21 C. Similarly, delivery of lucifer yellow into cells with poly-APS was enhanced at lower temperatures. Transfection with cDNA encoding for the expression enhanced green fluorescent protein was also evaluated at 12 C with poly-APS, lipofectamine and LipoGen. Intracellular delivery of siRNA was achieved with knockdown in beta-actin expression when lipofectamine and LipoGen were used as transfection reagents. However, intracellular delivery of siRNA was not achieved with poly-APS. Conclusion: Poly-APS mediated pore formation is critical to its activity as a transfection reagent, but lipofection systems utilise distinct mechanisms to enable delivery of DNA and siRNA into cells.

AB - Background: Haplosclerid marine sponges produce pore forming polyalkylpyridinium salts ( poly-APS), which can be used to deliver macromolecules into cells. The aim of this study was to investigate the delivery of DNA, siRNA and lucifer yellow into cells mediated by poly-APS and its potential mechanisms as compared with other lipofection systems ( lipofectamine and N-4, N(9-)dioleoylspermine (LipoGen)). DNA condensation was evaluated and HEK 293 and HtTA HeLa cells were used to investigate pore formation and intracellular delivery of cDNA, siRNA and lucifer yellow. Results: Poly-APS and LipoGen were both found to be highly efficient DNA condensing agents. Fura-2 calcium imaging was used to measure calcium transients indicative of cell membrane pore forming activity. Calcium transients were evoked by poly-APS but not LipoGen and lipofectamine. The increases in intracellular calcium produced by poly-APS showed temperature sensitivity with greater responses being observed at 12 C compared to 21 C. Similarly, delivery of lucifer yellow into cells with poly-APS was enhanced at lower temperatures. Transfection with cDNA encoding for the expression enhanced green fluorescent protein was also evaluated at 12 C with poly-APS, lipofectamine and LipoGen. Intracellular delivery of siRNA was achieved with knockdown in beta-actin expression when lipofectamine and LipoGen were used as transfection reagents. However, intracellular delivery of siRNA was not achieved with poly-APS. Conclusion: Poly-APS mediated pore formation is critical to its activity as a transfection reagent, but lipofection systems utilise distinct mechanisms to enable delivery of DNA and siRNA into cells.

KW - gene delivery

KW - Reniera-sarai

KW - membrane-properties

KW - Haliclona-vicosa

KW - ethidium-bromide

KW - mammalian-cells

KW - DNA

KW - spermine

KW - condensation

KW - transfection

U2 - 10.1186/1472-6750-6-6

DO - 10.1186/1472-6750-6-6

M3 - Article

VL - 6

JO - BMC Biotechnology

JF - BMC Biotechnology

SN - 1472-6750

M1 - 6

ER -