Free Energy Landscaping for Single-Molecule Biophysics

单分子生物物理学的自由能源景观

• 批准号: 1409577
• 负责人: Derek Stein
• 金额: $ 37.5万
• 依托单位: Brown University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1409577&HistoricalAwards=false
• 关键词: Free; Energy; Landscaping; Single; Molecule

Non-technical: This award by the Biomaterials program in the Division of Materials Research to Brown University is cofunded by the Instrument Development for Biological Research program in the Division of Biological Infrastructure (BIO). This award will study "Lab-on-a-chip" technology in improving healthcare by making biomedical tests smaller, cheaper, and faster. The ultimate limit would be tests on single molecules. This project aims to take an important step in that direction by developing powerful ways of controlling the shape and the motion of single DNA molecules. With this award, techniques will be developed that require nanofluidic device "channels with dimensions only tens to hundreds of nanometers across" because they can exploit physical concepts that only apply at that scale. For instance, a fluidic device with nanoscale topographic features inside it, like nanotrenches, harnesses the entropy of a DNA molecule to guide its motion or even contort it into predetermined shapes. Entropy is a measure of the multitude of microscopically different shapes a polymer can adopt. It is also possible to manipulate a DNA molecule electrostatically, using a nanofluidic version of the field effect upon which transistors are based. The experiments in this project will advance our fundamental understanding of nanofluidics and polymer physics. This project will also apply control over single DNA molecules to obtain overlapping restriction maps of a single molecule, which can increase the throughput of long-range sequence information that complements next-generation genomic sequencing data. This interdisciplinary project will prepare graduate and undergraduate students for careers in a rapidly growing area of the high-technology economy. Furthermore, the videos collected of single DNA molecules being controlled one by one will be used in the classroom at Brown University and in the nearby public elementary schools. Technical: This award is to develop new methods in controlling the configurations and the transport of single DNA molecules inside nanofluidic devices. The proposed methods rely on influencing the free energy landscape for a nanoconfined polyelectrolyte. This researcher will investigate how a series of trenches embedded within a nanoslit governs the configurational entropy of a polymer through the reduction of the number of configurations available in shallow regions as compared with deeper regions. The investigator will also explore how the charged inner surfaces of a device influence the enthalpy of a confined molecule through Coulomb forces, which can be locally tuned using electrodes. These new modalities of control should enable fundamental studies on single DNA molecules and new bio-analytical applications. Stochastic resonance and noise-assisted transport of DNA will be studied in a synthetic, bumpy free energy landscape. A similar device will be used to create overlapping restriction maps of the same DNA molecule by performing multiple restriction digests, sequentially, on the molecule as it is trapped and stretched in a long nanotrench. This project will advance "lab-on-a-chip" applications, provide excellent training opportunities for graduate and undergraduate students, and will support outreach activities to local public schools.

非技术性：生物材料计划在材料研究部门颁发的布朗大学的奖项是由生物基础设施（BIO）生物学仪器开发计划（BIO）的奖项。该奖项将通过使生物医学测试更小，更便宜和更快地研究“实验室芯片”技术，以改善医疗保健。最终的极限将是对单分子的测试。该项目旨在通过开发强大的方法来控制单个DNA分子的形状和运动来迈出重要的一步。通过此奖项，将开发需要纳米流体设备的技术“尺寸仅数十万到数百纳米的通道”，因为它们可以利用仅适用于该规模的物理概念。例如，内部具有纳米级地形特征的流体装置，例如纳米肾上腺，利用DNA分子的熵引导其运动，甚至扭曲其为预定的形状。熵是聚合物可以采用的多种显微镜不同形状的量度。也可以使用晶体管所基于的田间效应的纳米流体版本来操纵DNA分子。该项目中的实验将提高我们对纳米流体和聚合物物理学的基本理解。该项目还将对单个DNA分子进行控制，以获得单个分子的重叠限制图，这可以增加与下一代基因组测序数据相辅相成的远程序列信息的吞吐量。这个跨学科项目将为毕业生和本科生做好准备，以在高科技经济的快速发展领域的职业中做好准备。此外，收集的单个DNA分子的视频将一一控制在布朗大学的课堂和附近的公立小学中。技术：该奖项是为控制纳米流体设备内的单个DNA分子的构型和传输而开发新方法。所提出的方法依赖于纳米夹层聚电解质的自由能景观。该研究人员将研究一系列嵌入纳米斜率内的沟槽如何通过减少与更深层区域相比浅区域可用的配置数量来控制聚合物的构型熵。研究者还将探索设备的带电的内表面如何通过库仑力影响狭窄的分子的焓，库仑力可以使用电极局部调节。这些新的控制方式应使对单个DNA分子和新的生物分析应用的基本研究。随机共振和DNA的噪声辅助运输将在合成的，颠簸的自由能景观中进行研究。类似的设备将用于创建相同DNA分子的重叠限制图，通过在分子中依次在分子上进行多种限制消化，因为它被困在长纳米型中。该项目将推进“芯片实验室”的应用程序，为研究生和本科生提供极好的培训机会，并将支持向当地公立学校进行外展活动。