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Fluorescence Cross-Correlation Spectroscopy as a valuable tool to characterize cationic liposome-DNA nanoparticle assembly.
Journal of Biophotonics ( IF 2.0 ) Pub Date : 2020-08-21 , DOI: 10.1002/jbio.202000200
Ana I Gómez-Varela 1, 2 , Ricardo Gaspar 1 , Adelaide Miranda 1 , Juliane L Assis 3 , Rafael H F Valverde 3 , Marcelo Einicker-Lamas 3 , Bruno F B Silva 1 , Pieter A A De Beule 1
Affiliation  

The development of nonviral gene delivery vehicles for therapeutic applications requires methods capable of quantifying the association between the genes and their carrier counterparts. Here we investigate the potential of fluorescence cross‐correlation spectroscopy (FCCS) to characterize and optimize the assembly of nonviral cationic liposome (CL)‐DNA complexes based on a CL formulation consisting of the cationic lipid DOTAP and zwitterionic lipid DOPC. We use a DNA plasmid for lipoplex loading encoding the Oct4 gene, critically involved in reprogramming somatic cells into induced pluripotent stem cells. We demonstrate that FCCS is able to quantitatively determine the extent of the association between DNA and the liposomes and assess its loading capacity. We also establish that the cationic lipid fraction, being proportional to the liposome membrane charge density, as well as charge ratio between the CLs and anionic DNA play an important role in the degree of interaction between the liposomes and DNA.image

中文翻译:

荧光互相关光谱法是表征阳离子脂质体-DNA纳米粒子组装的有价值的工具。

用于治疗应用的非病毒基因递送载体的开发需要能够定量基因与其载体对应物之间关联的方法。在这里我们研究了荧光互相关光谱(FCCS)表征和优化基于阳离子脂质DOTAP和两性离子脂质DOPC的CL配方的非病毒阳离子脂质体(CL)-DNA复合物组装的潜力。我们使用DNA质粒进行lipoplex加载,编码Oct4基因,关键参与将体细胞重编程为诱导性多能干细胞。我们证明,FCCS能够定量确定DNA与脂质体之间的缔合程度并评估其负载能力。我们还确定了阳离子脂质部分图片
更新日期:2020-08-21
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