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Tuning Interfacial Biomolecule Interactions with Massively Parallel Nanopore Arrays

使用大规模并行纳米孔阵列调节界面生物分子相互作用

基本信息

批准号：
1704901
负责人：
Matthew Kipper
金额：
$ 41.09万
依托单位：
Colorado State University
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-15 至 2021-10-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1704901&HistoricalAwards=false
关键词：
Tuning Interfacial Biomolecule Interactions Massively

项目摘要

DNA stores biological information with a very high density. New tools of genomics and genetic engineering that have emerged over the past 20 years, providing the transformative capabilities to both "read" DNA, and to "write" DNA. New biosensors that diagnose diseases and predict responses to treatments can be based on "reading" DNA, and development of new pest-resistant crops, for example, can be based on "writing" DNA. While DNA is protected when inside living cells, it is relatively unstable when exposed to environments outside of cells. This work will advance our understanding of how DNA interacts with a new class of materials with DNA-sized, nanoscale pores. The chemistry of these pores can be tuned to optimize interactions with DNA, for storage, stability, and for reporting specific chemical binding events. In future work, these materials could be used to develop new biosensors and advanced nanomaterials that could store DNA or transduce mechanical and chemical signals through controlled DNA binding. We will study the interactions of short DNA segments with the surfaces and pores of crosslinked protein crystals, by atomic force microscopy and adsorption isotherm measurements. Diffusion of DNA within nanopores will be characterized by fluorescence microscopy methods. Finally, we will assess the ability of protein crystals to: stabilize guest DNA, couple DNA-pore interactions to ligand binding on the protein crystal surface, and report DNA hybridization through fluorescence.

DNA以非常高的密度存储生物信息。在过去20年中出现的基因组学和基因工程的新工具，提供了“读取”DNA和“写入”DNA的变革能力。诊断疾病和预测治疗反应的新生物传感器可以基于“阅读”DNA，例如，新的抗虫害作物的开发可以基于“写入”DNA。虽然DNA在活细胞内受到保护，但当暴露于细胞外环境时相对不稳定。这项工作将推进我们对DNA如何与一类具有DNA大小的纳米级孔的新材料相互作用的理解。可以调整这些孔的化学性质以优化与DNA的相互作用，用于储存、稳定性和报告特定的化学结合事件。在未来的工作中，这些材料可用于开发新的生物传感器和先进的纳米材料，这些材料可以通过控制DNA结合来存储DNA或存储机械和化学信号。我们将通过原子力显微镜和吸附等温线测量来研究短DNA片段与交联蛋白质晶体的表面和孔的相互作用。DNA在纳米孔内的扩散将通过荧光显微镜方法表征。最后，我们将评估蛋白质晶体的能力：稳定客体DNA，耦合DNA-孔相互作用的蛋白质晶体表面上的配体结合，并通过荧光报告DNA杂交。