TY - JOUR

T1 - DNA-polycation complexes Effect of polycation structure on physico-chemical and biological properties

AU - Slita, A. V.

AU - Kasyanenko, N. A.

AU - Nazarova, O. V.

AU - Gavrilova, I. I.

AU - Eropkina, E. M.

AU - Sirotkin, A. K.

AU - Smirnova, T. D.

AU - Kiselev, O. I.

AU - Panarin, E. F.

N1 - Funding Information: The authors thank Dr. V.V. Zarubaev for valuable discussion and the Ministry of Education and Science of Russia for financial support (grant NSh 1823.2003.3). Copyright: Copyright 2008 Elsevier B.V., All rights reserved.

PY - 2007/1/20

Y1 - 2007/1/20

N2 - The purpose of the study was to investigate the influence of cationic polymer structure on the formation of DNA-polycation complexes and their transfection activity. Primary, tertiary, and quaternary polyamines with molecular masses ranging from 8000 to 200,000 were investigated. DNA-cationic polymer interaction was characterized by low gradient viscometry, dynamic light scattering, circular dichroism, UV spectrometry, flow birefringence, DNA electrophoresis, and electron microscopy. Transfection activity of the complexes was evaluated by the expression of reporter gene (β-galactosidase) and using synthetic FITC-labelled oligonucleotides. Complex formation was found to be dependent on the structure and molecular weight of the polymer and the ionic strength of the solution. Secondary DNA structure in complexes was not disrupted, and DNA was protected from protonation. Cell lines of different origin were used for testing of transfection activity of the complexes. The sensitivity of the cells to transfection was established to be highly dependent on the cell line. DNA-polycation complexes are non-toxic according to MTT. Polyallylamine, and polydimethylaminoethylmethacrylate were found to be the most promising polycations for gene delivery. Transfection efficacy of their complexes with DNA to T-98G cells reaches up to 90-100%. It was found that optimal molecular mass of polydimethylaminoethylmethacrylate is in the range of 8000-50,000 Da.

AB - The purpose of the study was to investigate the influence of cationic polymer structure on the formation of DNA-polycation complexes and their transfection activity. Primary, tertiary, and quaternary polyamines with molecular masses ranging from 8000 to 200,000 were investigated. DNA-cationic polymer interaction was characterized by low gradient viscometry, dynamic light scattering, circular dichroism, UV spectrometry, flow birefringence, DNA electrophoresis, and electron microscopy. Transfection activity of the complexes was evaluated by the expression of reporter gene (β-galactosidase) and using synthetic FITC-labelled oligonucleotides. Complex formation was found to be dependent on the structure and molecular weight of the polymer and the ionic strength of the solution. Secondary DNA structure in complexes was not disrupted, and DNA was protected from protonation. Cell lines of different origin were used for testing of transfection activity of the complexes. The sensitivity of the cells to transfection was established to be highly dependent on the cell line. DNA-polycation complexes are non-toxic according to MTT. Polyallylamine, and polydimethylaminoethylmethacrylate were found to be the most promising polycations for gene delivery. Transfection efficacy of their complexes with DNA to T-98G cells reaches up to 90-100%. It was found that optimal molecular mass of polydimethylaminoethylmethacrylate is in the range of 8000-50,000 Da.

KW - Polyallylamine

KW - Polycation-DNA complexes

KW - Polydimethylaminoethylmethacrylate

KW - Transfection activity

UR - http://www.scopus.com/inward/record.url?scp=33845633265&partnerID=8YFLogxK

U2 - 10.1016/j.jbiotec.2006.07.016

DO - 10.1016/j.jbiotec.2006.07.016

M3 - Article

C2 - 16934901

AN - SCOPUS:33845633265

VL - 127

SP - 679

EP - 693

JO - Journal of Biotechnology

JF - Journal of Biotechnology

SN - 0168-1656

IS - 4

ER -