DNA–polycation complexes Effect of polycation structure on physico-chemical and biological properties
- 作者：A.V. Slita ; N.A. Kasyanenko ; O.V. Nazarova ; I.I. Gavrilova ; E.M. Eropkina ; A.K. Sirotkin ; T.D. Smirnova ; O.I. Kiselev ; E.F. Panarin
- 关键词：Polycation– ; DNA complexes ; Polyallylamine ; Polydimethylaminoethylmethacrylate ; Transfection activity
- 刊名：Journal of Biotechnology
- 出版年：2007
- 期刊代码：53_01681656
- 类别：imm
- 出版时间：20 January 2007
- 卷：127
- 期：4
- 页码：679-693
- 文件大小：1367 K
摘要
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.