NIRT: Manipulation of DNA-Protein Interactions at the Nano-Scale

0304316 Larson该提案是对纳米科学与工程倡议（NSF 02-148，类别NIRT）的响应。本研究的目的是设计和建立一个微加工平台，用于研究纳米尺度下的DNA/蛋白质相互作用。这些相互作用是维持和转录DNA的细胞机制的核心，包括转录因子以及组蛋白和其他染色质蛋白。这些DNA/蛋白质相互作用正在细胞水平上进行积极的研究，但对它们进行详细了解的下一步需要在更小的长度尺度上进行研究，包括纳米尺度。因此，将构建微制造装置，其包含长度为20至30微米且宽度范围为20 nm至2微米的主通道。单个DNA分子将在通道中拉伸并使用高频电场定位，并使用化学连接到DNA分子末端的硫醇基团锚定在金电极上。这个长的、非常薄的通道将连接到薄的侧通道，用于将DNA相互作用蛋白局部添加到DNA分子的特定区域。蛋白质将通过使用捕获电极以及使用流过其它侧流的流体动力学聚焦和其它限制方法进一步定位。通道将在硅中蚀刻，并结合到薄玻璃基板上，以允许通过光学显微镜进行检查。该提案的更广泛影响包括将拟议的研究与为研究生引入新的纳米科学与工程（NSE）课程相结合。新课程将集中在与微流体电子功能集成相关的纳米制造问题和与生物样品处理相关的运输问题。

0304316LarsonThis proposal was received in response to Nanoscale Science and Engineering initiative, NSF 02-148, category NIRT. The objective of this proposal is to design and build a microfabricated platform for study of DNA/protein interactions at the nano-scale. These interactions are at the heart of the cellular machinery for maintaining and transcribing DNA, and include transcription factors, as well as histones and other chromatin proteins. These DNA/protein interactions are under active study at the cellular level, but the next steps towards a detailed understanding of them require their study at smaller length scales, including the nanoscale. Therefore, a microfabricated devise will be built, which contains a main channel of length 20 to 30 microns and of width ranging from 20 nm to 2 microns. Individual DNA molecules will be stretched in the channel and positioned using high frequency electric fields, and anchored at gold electrodes using thiol groups chemically attached to the ends of the DNA molecules. This long, very thin, channel will be joined to thin side channels for addition of DNA-interacting proteins locally to specific regions of the DNA molecule. The proteins will be further localized by use of capture electrodes as well as hydrodynamic focusing using flow through other side streams, and other confinement methods. The channels will be etched in silicon, and bonded to a thin glass substrate, to permit examination by optical microscopy. The broader impacts of the proposal include the integration of the proposed research with the introduction of a new Nanoscale Science and Engineering (NSE) curriculum for graduate students. The new course will focus on nano- fabrication issues related to integration of electronic function with microfluidics and transport issues related to the processing of biological samples.