Phosphonium-containing polyelectrolytes for nonviral gene delivery

Sean T. Hemp, Michael H. Allen, Matthew D. Green, Timothy Edward Long

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

Nonviral gene therapy focuses intensely on nitrogen-containing macromolecules and lipids to condense and deliver DNA as a therapeutic for genetic human diseases. For the first time, DNA binding and gene transfection experiments compared phosphonium-containing macromolecules with their respective ammonium analogs. Conventional free radical polymerization of quaternized 4-vinylbenzyl chloride monomers afforded phosphonium- and ammonium-containing homopolymers for gene transfection experiments of HeLa cells. Aqueous size exclusion chromatography confirmed similar absolute molecular weights for all polyelectrolytes. DNA gel shift assays and luciferase expression assays revealed phosphonium-containing polymers bound DNA at lower charge ratios and displayed improved luciferase expression relative to the ammonium analogs. The triethyl-based vectors for both cations failed to transfect HeLa cells, whereas tributyl-based vectors successfully transfected HeLa cells similar to Superfect demonstrating the influence of the alkyl substituent lengths on the efficacy of the gene delivery vehicle. Cellular uptake of Cy5-labeled DNA highlighted successful cellular uptake of triethyl-based polyplexes, showing that intracellular mechanisms presumably prevented luciferase expression. Endocytic inhibition studies using genistein, methyl β-cyclodextrin, or amantadine demonstrated the caveolae-mediated pathway as the preferred cellular uptake mechanism for the delivery vehicles examined. Our studies demonstrated that changing the polymeric cation from ammonium to phosphonium enables an unexplored array of synthetic vectors for enhanced DNA binding and transfection that may transform the field of nonviral gene delivery.

Original languageEnglish
Pages (from-to)231-238
Number of pages8
JournalBiomacromolecules
Volume13
Issue number1
DOIs
Publication statusPublished - 2012 Jan 9
Externally publishedYes

Fingerprint

Polyelectrolytes
DNA
Genes
Ammonium Compounds
Luciferases
Macromolecules
Cations
Assays
Positive ions
Amantadine
Gene therapy
Size exclusion chromatography
Genistein
Cyclodextrins
Free radical polymerization
Homopolymerization
Lipids
Chlorides
Polymers
Nitrogen

ASJC Scopus subject areas

  • Bioengineering
  • Materials Chemistry
  • Polymers and Plastics
  • Biomaterials

Cite this

Hemp, S. T., Allen, M. H., Green, M. D., & Long, T. E. (2012). Phosphonium-containing polyelectrolytes for nonviral gene delivery. Biomacromolecules, 13(1), 231-238. https://doi.org/10.1021/bm201503a

Phosphonium-containing polyelectrolytes for nonviral gene delivery. / Hemp, Sean T.; Allen, Michael H.; Green, Matthew D.; Long, Timothy Edward.

In: Biomacromolecules, Vol. 13, No. 1, 09.01.2012, p. 231-238.

Research output: Contribution to journalArticle

Hemp, ST, Allen, MH, Green, MD & Long, TE 2012, 'Phosphonium-containing polyelectrolytes for nonviral gene delivery', Biomacromolecules, vol. 13, no. 1, pp. 231-238. https://doi.org/10.1021/bm201503a
Hemp, Sean T. ; Allen, Michael H. ; Green, Matthew D. ; Long, Timothy Edward. / Phosphonium-containing polyelectrolytes for nonviral gene delivery. In: Biomacromolecules. 2012 ; Vol. 13, No. 1. pp. 231-238.
@article{6d549312c5b74713a6d9ef1d81a6bd0b,
title = "Phosphonium-containing polyelectrolytes for nonviral gene delivery",
abstract = "Nonviral gene therapy focuses intensely on nitrogen-containing macromolecules and lipids to condense and deliver DNA as a therapeutic for genetic human diseases. For the first time, DNA binding and gene transfection experiments compared phosphonium-containing macromolecules with their respective ammonium analogs. Conventional free radical polymerization of quaternized 4-vinylbenzyl chloride monomers afforded phosphonium- and ammonium-containing homopolymers for gene transfection experiments of HeLa cells. Aqueous size exclusion chromatography confirmed similar absolute molecular weights for all polyelectrolytes. DNA gel shift assays and luciferase expression assays revealed phosphonium-containing polymers bound DNA at lower charge ratios and displayed improved luciferase expression relative to the ammonium analogs. The triethyl-based vectors for both cations failed to transfect HeLa cells, whereas tributyl-based vectors successfully transfected HeLa cells similar to Superfect demonstrating the influence of the alkyl substituent lengths on the efficacy of the gene delivery vehicle. Cellular uptake of Cy5-labeled DNA highlighted successful cellular uptake of triethyl-based polyplexes, showing that intracellular mechanisms presumably prevented luciferase expression. Endocytic inhibition studies using genistein, methyl β-cyclodextrin, or amantadine demonstrated the caveolae-mediated pathway as the preferred cellular uptake mechanism for the delivery vehicles examined. Our studies demonstrated that changing the polymeric cation from ammonium to phosphonium enables an unexplored array of synthetic vectors for enhanced DNA binding and transfection that may transform the field of nonviral gene delivery.",
author = "Hemp, {Sean T.} and Allen, {Michael H.} and Green, {Matthew D.} and Long, {Timothy Edward}",
year = "2012",
month = "1",
day = "9",
doi = "10.1021/bm201503a",
language = "English",
volume = "13",
pages = "231--238",
journal = "Biomacromolecules",
issn = "1525-7797",
publisher = "American Chemical Society",
number = "1",

}

TY - JOUR

T1 - Phosphonium-containing polyelectrolytes for nonviral gene delivery

AU - Hemp, Sean T.

AU - Allen, Michael H.

AU - Green, Matthew D.

AU - Long, Timothy Edward

PY - 2012/1/9

Y1 - 2012/1/9

N2 - Nonviral gene therapy focuses intensely on nitrogen-containing macromolecules and lipids to condense and deliver DNA as a therapeutic for genetic human diseases. For the first time, DNA binding and gene transfection experiments compared phosphonium-containing macromolecules with their respective ammonium analogs. Conventional free radical polymerization of quaternized 4-vinylbenzyl chloride monomers afforded phosphonium- and ammonium-containing homopolymers for gene transfection experiments of HeLa cells. Aqueous size exclusion chromatography confirmed similar absolute molecular weights for all polyelectrolytes. DNA gel shift assays and luciferase expression assays revealed phosphonium-containing polymers bound DNA at lower charge ratios and displayed improved luciferase expression relative to the ammonium analogs. The triethyl-based vectors for both cations failed to transfect HeLa cells, whereas tributyl-based vectors successfully transfected HeLa cells similar to Superfect demonstrating the influence of the alkyl substituent lengths on the efficacy of the gene delivery vehicle. Cellular uptake of Cy5-labeled DNA highlighted successful cellular uptake of triethyl-based polyplexes, showing that intracellular mechanisms presumably prevented luciferase expression. Endocytic inhibition studies using genistein, methyl β-cyclodextrin, or amantadine demonstrated the caveolae-mediated pathway as the preferred cellular uptake mechanism for the delivery vehicles examined. Our studies demonstrated that changing the polymeric cation from ammonium to phosphonium enables an unexplored array of synthetic vectors for enhanced DNA binding and transfection that may transform the field of nonviral gene delivery.

AB - Nonviral gene therapy focuses intensely on nitrogen-containing macromolecules and lipids to condense and deliver DNA as a therapeutic for genetic human diseases. For the first time, DNA binding and gene transfection experiments compared phosphonium-containing macromolecules with their respective ammonium analogs. Conventional free radical polymerization of quaternized 4-vinylbenzyl chloride monomers afforded phosphonium- and ammonium-containing homopolymers for gene transfection experiments of HeLa cells. Aqueous size exclusion chromatography confirmed similar absolute molecular weights for all polyelectrolytes. DNA gel shift assays and luciferase expression assays revealed phosphonium-containing polymers bound DNA at lower charge ratios and displayed improved luciferase expression relative to the ammonium analogs. The triethyl-based vectors for both cations failed to transfect HeLa cells, whereas tributyl-based vectors successfully transfected HeLa cells similar to Superfect demonstrating the influence of the alkyl substituent lengths on the efficacy of the gene delivery vehicle. Cellular uptake of Cy5-labeled DNA highlighted successful cellular uptake of triethyl-based polyplexes, showing that intracellular mechanisms presumably prevented luciferase expression. Endocytic inhibition studies using genistein, methyl β-cyclodextrin, or amantadine demonstrated the caveolae-mediated pathway as the preferred cellular uptake mechanism for the delivery vehicles examined. Our studies demonstrated that changing the polymeric cation from ammonium to phosphonium enables an unexplored array of synthetic vectors for enhanced DNA binding and transfection that may transform the field of nonviral gene delivery.

UR - http://www.scopus.com/inward/record.url?scp=84855605822&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84855605822&partnerID=8YFLogxK

U2 - 10.1021/bm201503a

DO - 10.1021/bm201503a

M3 - Article

C2 - 22136386

AN - SCOPUS:84855605822

VL - 13

SP - 231

EP - 238

JO - Biomacromolecules

JF - Biomacromolecules

SN - 1525-7797

IS - 1

ER -