Shape Control and Transfection of Self-assembled Polymer-DNA Nanoparticles

描述（由申请人提供）：本研究的总体目标是从机理上理解自组装DNA纳米颗粒的形状控制，并检验纳米颗粒形状可以影响其细胞摄取、细胞内运输和基因递送效率的假设。最近的一些研究提出了前景，即病毒颗粒和各种类型的合成纳米颗粒的形态是其运输性质和生物功能的重要决定因素。我们已经开发了一种方法，用于自组装的DNA含有纳米粒子具有几个不同的形状（球形，棒状和蠕虫状）类似于一些病毒颗粒的基因转染的目的。这样的纳米颗粒是用于理解DNA诱导的自组装的机制以及纳米颗粒形状对其稳定性、细胞-纳米颗粒相互作用、转染效率和体内转运动力学的影响的理想系统。通过这项探索性资助，我们计划（1）确定有效控制纳米颗粒形状和尺寸的关键实验参数，并使用实验和计算建模相结合的方法来理解聚乙二醇化聚阳离子在DNA缩合中的形状控制机制;和（2）为了证明细胞摄取中的纳米颗粒形状依赖性，在肝靶向基因递送模型中的体外和体内细胞内运输和转染效率。这项研究将提供一个主要的驱动力的DNA/PEG-聚阳离子纳米粒子的自组装机制的理解，并确定其形状控制所涉及的关键参数。这将提供一种有效的方法来控制DNA/聚合物纳米颗粒的大小和形状，可以适用于各种聚乙二醇化的基因载体，在合成纳米颗粒的高度控制其形状或形态。

DESCRIPTION (provided by applicant): The overall objective of this study is to develop a mechanistic understanding of shape control of self-assembled DNA nanoparticles, and to test the hypothesis that nanoparticle shape can influence their cellular uptake, intracellular trafficking and gene delivery efficiency. Several recent studies have raised the prospect that the morphology of virus particles and various types of synthetic nanoparticles is an important determinant for their transport properties and biological functions. We have developed a method for the self-assembly of DNA-containing nanoparticles with several distinct shapes (spherical, rod-like and worm-like) similar to some viral particles for the purpose of gene transfection. Such nanoparticles are ideal systems for understanding the mechanism of DNA-induced self-assembly and the effect of nanoparticle shape on their stability, cell- nanoparticle interactions, transfection efficiency and in vivo transport kinetics. With this Exploratory Grant, we plan (1) to determine the key experimental parameters that effectively control the shape and size of nanoparticles, and to understand the mechanism of shape control in DNA condensation by PEGylated polycations using a combined experimental and computational modeling approach; and (2) to demonstrate nanoparticle shape dependence in cellular uptake, intracellular trafficking and transfection efficiency in vitro and in vivo in a liver-targeted gene delivery model. This study will provide a mechanistic understanding of the major driving forces for the self-assembly of DNA/PEG-polycation nanoparticles and identify key parameters involved in their shape control. It will offer an effective method to control the size and shape of DNA/polymer nanoparticles that can be applicable to a variety of PEGylated gene carriers in synthesizing nanoparticles with high degree of control over their shapes or morphologies.