Nanoparticles with Tailored Monolayers for DNA Transfection

用于 DNA 转染的定制单层纳米颗粒

基本信息

批准号：
7140469
负责人：
VINCENT M. ROTELLO
金额：
$ 22.3万
依托单位：
UNIVERSITY OF MASSACHUSETTS AMHERST
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-08-15 至 2008-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7140469
关键词：
DNA biotechnology confocal scanning microscopy gene delivery system intracellular transport molecular film nanotechnology small molecule technology /technique development transfection /expression vector

项目摘要

DESCRIPTION (provided by applicant): Nucleic acid transfection is one of the most important tools in modern biology, and an emerging tool for biomedical applications. In our proposed research, we will use gold nanoparticles as transfection vectors. Gold nanoparticles possess three attributes that make them highly promising vectors: 1) Tunable size commensurate with biomolecular scales (4-10 nm); 2) Low inherent toxicity; 3) Stable and easily decorated monolayer surface, allowing control over surface charge and hydrophobicity, as well as facile conjugation to biotags such as peptides, and other functional molecules such as fluorescent dyes. Our proposed research features three specific Aims. These Aims are interrelated and will be performed concurrently, bringing to bear our expertise in synthesis, nanomaterials, and biology. In Aim 1, we will systematically vary the size of the particle and the structure of the monolayer. These studies will be used to create optimized transfection vectors, as well as exploring new vector-DNA unpackaging strategies. We will also explore the conjugation of peptide tags to the particles to provide enhanced uptake and localization of the DNA-MMPC aggregates. Aim 2 describes the strategies that will be used to quantify particle-DNA interactions, transfection efficiency, and toxicity. Aim 3 describes the use of microscopy to determine bottlenecks in the transfection process, providing information that will be used to design enhanced vectors. Rationale for R21 Mechanism. The major innovation of this proposal is the synergistic application of nanotechnology and synthesis to the area of transfection. The proposed research uses the gold core as a scaffold for the presentation of diverse functionalities, providing systems that should provide high transfection coupled with low toxicity. The R21 grant will provide an opportunity to explore the potentialities, directly providing enhanced vectors for in vitro studies. These studies will also lay the groundwork for the development of in vivo gene delivery systems that will be pursued in future studies.

描述（由申请人提供）：核酸转染是现代生物学中最重要的工具之一，也是生物医学应用的新兴工具。在我们提出的研究中，我们将使用金纳米颗粒作为转染载体。金纳米颗粒具有三个属性，使其成为非常有前途的载体：1）与生物分子尺度相称的可调尺寸（4-10 nm）； 2）固有毒性低； 3) 稳定且易于装饰的单层表面，可以控制表面电荷和疏水性，以及轻松与生物标签（如肽）和其他功能分子（如荧光染料）缀合。我们提出的研究具有三个具体目标。这些目标是相互关联的，并将同时实现，发挥我们在合成、纳米材料和生物学方面的专业知识。在目标 1 中，我们将系统地改变颗粒的尺寸和单层的结构。这些研究将用于创建优化的转染载体，以及探索新的载体 DNA 解包装策略。我们还将探索肽标签与颗粒的结合，以增强 DNA-MMPC 聚集体的吸收和定位。目标 2 描述了用于量化颗粒-DNA 相互作用、转染效率和毒性的策略。目标 3 描述了使用显微镜来确定转染过程中的瓶颈，提供用于设计增强载体的信息。 R21 机制的基本原理。该提案的主要创新在于纳米技术和合成在转染领域的协同应用。拟议的研究使用金核心作为呈现多种功能的支架，提供应提供高转染和低毒性的系统。 R21 资助将提供一个探索潜力的机会，直接为体外研究提供增强的载体。这些研究还将为未来研究中的体内基因传递系统的开发奠定基础。