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Published online 2 January 2004

Nucleic Acids Research, 2004, Vol. 32, No. 1 127-134
© 2004 Oxford University Press

Formation of DNA nanoparticles in the presence of novel polyamine analogues: a laser light scattering and atomic force microscopic study

Veena Vijayanathan1, Thresia Thomas2,3,4, Thomas Antony5, Akira Shirahata6 and T. J. Thomas*,1,3

Departments of 1 Medicine, and 2 Environmental and Community Medicine, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, 125 Paterson Street, CAB 7090, New Brunswick, NJ 08903, 3 The Cancer Institute of New Jersey, New Brunswick, NJ 08903, 4 Environmental and Occupational Health Sciences Institute, Piscataway, NJ 08854, USA, 5 Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, D-37077, Göttingen, Germany and 6 Department of Biochemistry and Cellular Physiology, Josai University, Saitama, Japan

*To whom correspondence should be addressed. Tel: +1 732 235 8460; Fax: +1 732 235 8473; Email: thomastj{at}UMDNJ.edu

We synthesized a pentamine (3-3-3-3) and two hexamine (3-3-3-3-3 and 3-4-3-4-3) analogues of the natural polyamine, spermine (3-4-3) and studied their effectiveness in condensing pGL3 plasmid DNA, using light scattering and atomic force microscopic (AFM) techniques. The midpoint concentration of the polyamines on pGL3 condensation (EC50) was 11.3, 10.6, 1.5, 0.49 and 0.52 µM, respectively, for 3-4-3, norspermine (3-3-3), 3-3-3-3, 3-3-3-3-3 and 3-4-3-4-3 in 10 mM Na cacodylate buffer. Dynamic laser light scattering study showed a decrease in hydrodynamic radii of plasmid DNA particles as the number of positive charges on the polyamines increased. AFM data showed the presence of toroids with outer diameter of 117–191 nm for different polyamines, and a mean height of 2.61 ± 0.77 nm. AFM results also revealed the presence of intermediate structures, including those showing circumferential winding of DNA to toroids. The dependence of the EC50 on Na+ concentration suggests different modes of binding of spermine and its higher valent analogues with DNA. Our results show a 20-fold increase in the efficacy of hexamines for DNA condensation compared to spermine, and provide new insights into the mechanism(s) of DNA nanoparticle formation. These studies might help to develop novel nonviral gene delivery vehicles.


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