Reitan Nina K, Maurstad Gjertrud, de Lange Davies Catharina, Strand Sabina P
Department of Physics, The Norwegian University of Science and Technology, Trondheim, Norway.
Biomacromolecules. 2009 Jun 8;10(6):1508-15. doi: 10.1021/bm900102d.
Chitosan can be used as a nonviral gene delivery vector for which DNA condensation and transfection efficacy strongly depend on structural parameters. In this study, we characterized the condensation of DNA by three molecularly tailored chitosans, including linear, trisaccharide substituted-, and self-branched trisaccharide substituted chitosan oligomers. No significant differences could be detected in the hydrodynamic diameters formed by the various chitosans as analyzed by dynamic light scattering. However, atomic force microscopy revealed that self-branched chitosan formed complexes with a higher ratio of globules to rods, and the heights of both globules and rods were larger than for complexes formed by the other chitosans. Using an amino/phosphate ratio of 10, fluorescence correlation spectroscopy measurements showed that self-branched chitosan exhibited a lower fraction (30%) of bound chitosan than the other chitosans. YOYO-1 was a superior fluorescent DNA-label compared to Cy5 and PicoGreen, since labeling with YOYO-1 had least effect on the size and structure of the complexes.
壳聚糖可作为一种非病毒基因传递载体，其DNA凝聚和转染效率在很大程度上取决于结构参数。在本研究中，我们对三种分子定制的壳聚糖（包括线性、三糖取代和自支化三糖取代壳聚糖低聚物）介导的DNA凝聚进行了表征。通过动态光散射分析发现，各种壳聚糖形成的流体动力学直径没有显著差异。然而，原子力显微镜显示，自支化壳聚糖形成的复合物中球状体与棒状体的比例更高，且球状体和棒状体的高度均大于其他壳聚糖形成的复合物。当氨基/磷酸比为10时，荧光相关光谱测量表明，自支化壳聚糖与其他壳聚糖相比，结合壳聚糖的比例更低（30%）。与Cy5和PicoGreen相比，YOYO-1是一种更优异的荧光DNA标记物，因为用YOYO-1标记对复合物的大小和结构影响最小。
Zotero 插件