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壳聚糖化合价对 DNA 络合和转染的影响:DNA-壳聚糖结合越弱,转染效率越高。

The influence of chitosan valence on the complexation and transfection of DNA: the weaker the DNA-chitosan binding the higher the transfection efficiency.

机构信息

Grupo de Nanomateriales y Materia Blanda, Departamento de Física de Materia Condensada, Facultad de Física, Universidad de Santiago de Compostela, E-15782 Santiago de Compostela, Spain.

出版信息

Colloids Surf B Biointerfaces. 2011 Jan 1;82(1):54-62. doi: 10.1016/j.colsurfb.2010.08.013. Epub 2010 Aug 14.

Abstract

The DNA-chitosan polyplexes have attracted for some years now the attention of physical-chemists and biologists for their potential use in gene therapy, however, the correlation between the physicochemical properties of these polyplexes with their transfection efficiency remains still unclear. In a recent paper we demonstrated by means of DLS that the DNA-chitosan complexation is favored at acidic conditions considering that fewer amounts of chitosan were required to compact the DNA. As a second study, in the present work we analyze the influence of chitosan valence on the complexation and transfection of DNA. Three chitosans of different molecular weights (three different valences) are characterized as gene carriers at 25°C and pH 5 over a wide range of chitosan-Nitrogen to DNA-Phosphate molar ratios, N/P, by means of conductometry, electrophoretic mobility, isothermal titration calorimetry (ITC), transmission electron microscopy (TEM), atomic force microscopy (AFM), and β-galactosidase and luciferase expression assays.

摘要

DNA-壳聚糖聚集体近年来引起了物理化学家与生物学家的关注,因为它们可能在基因治疗中得到应用。然而,这些聚集体的理化性质与转染效率之间的相关性仍然不清楚。在最近的一篇论文中,我们通过 DLS 证明,考虑到需要较少的壳聚糖来压缩 DNA,在酸性条件下有利于 DNA-壳聚糖的复合。作为第二项研究,在本工作中我们分析了壳聚糖化合价对 DNA 复合和转染的影响。通过电导率、电泳迁移率、等温滴定量热法 (ITC)、透射电子显微镜 (TEM)、原子力显微镜 (AFM) 以及β-半乳糖苷酶和荧光素酶表达测定,在广泛的壳聚糖-氮与 DNA-磷摩尔比(N/P)范围内,在 25°C 和 pH5 下,将三种不同分子量(三种不同化合价)的壳聚糖表征为基因载体。

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