Cationic lipids with increased DNA binding affinity for nonviral gene transfer in dividing and nondividing cells

Ajit S. Narang, Laura Thoma, Duane Miller, Ram I. Mahato

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Effect of headgroup structure on catonic lipid-mediated transfection was investigated with either a (i) tertiary amine, (ii) quaternary amine with a hydroxyl, or (iii) quaternary amine with mesylate as headgroups. Liposomes were formulated using cholesterol or dioleoyl phosphatidyl ethanolamine (DOPE) as colipids, and transfection efficiencies were determined in rapidly dividing colon carcinoma (CT 26) and rat aortic smooth muscle (RASM) cells as well as in nondividing human pancreatic islets using luciferase and green fluorescent protein expression plasmids, pcDNA3-Luc and pCMS-EGFP, respectively. Liposome/pDNA complexes were evaluated for DNA conformational state by circular dichroism (CD), DNA condensation by electrophoretic mobility shift assay (EMSA), particle size and zeta potential by laser diffraction technique, and surface morphology by transmission electron microscopy (TEM). Encouraging transfection results were obtained with the mesylate headgroup based lipid in liposome formulations with DOPE as a colipid, which were higher than the commercially available Lipofectamine formulation. We hypothesize that the additional hydrogen bonding or covalent interactions of the headgroup with the plasmid DNA, leading to higher binding affinity of the cationic lipids to pDNA, results in higher transfection. This hypothesis is supported by TEM observations where elongated complexes were observed and more lipid was seen associated with the DNA.

Original languageEnglish (US)
Pages (from-to)156-168
Number of pages13
JournalBioconjugate Chemistry
Volume16
Issue number1
DOIs
StatePublished - Jan 1 2005

Fingerprint

Gene transfer
Lipids
Transfection
Liposomes
DNA
Ethanolamines
Amines
Mesylates
Ethanolamine
Transmission Electron Microscopy
Genes
Plasmids
Transmission electron microscopy
Circular DNA
Electrophoretic mobility
Cholesterol
Dichroism
Electrophoretic Mobility Shift Assay
Zeta potential
Hydrogen Bonding

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

Cite this

Cationic lipids with increased DNA binding affinity for nonviral gene transfer in dividing and nondividing cells. / Narang, Ajit S.; Thoma, Laura; Miller, Duane; Mahato, Ram I.

In: Bioconjugate Chemistry, Vol. 16, No. 1, 01.01.2005, p. 156-168.

Research output: Contribution to journalArticle

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