Optical Single Channel Recording for Nanopore Sensing

用于纳米孔传感的光学单通道记录

• 批准号: BB/R001790/1
• 负责人: Mark Wallace
• 金额: $ 56.26万
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR001790%2F1
• 关键词: Optical; Single; Channel; Recording; Nanopore

Genome sequencing methods have led to a rapid expansion in our understanding of the subtleties and variations in how genetic profiles affect disease. This has been spurred in part by new technological advances. Nanopore sequencing has been highlighted as one upcoming technology with the potential to go further, and improve the speed and cost effectiveness of seqencing. In nanopore sensing, the flow of ions through a nanoscopic protein or solid-state pore is disrupted by the presence of an analyte. If this analyte is DNA, blockades in the current corresponding to the sequence are detected. Long reads of many thousands of bases can be made without amplification or labelling.Our research track record marks a long-standing interest in developing methods capable of detecting individual molecules using fluorescence microscopy. We have used this to improve our understanding of membrane protein function, and in particular, how specialized protein pores work. Most recently we used these methods to demonstrate DNA base detection in a nanopore using an optical, rather than an electrical readout. These methods have the potential to improve the parallelization of nanopore sensing.In collaboration with Oxford Nanopore Technologies we have identified two essential challenges for current nanopore sensing where an optical readout provides significant impact: 1. We will combine optical single channel recording and single-molecule fluorescence imaging to understand and then optimise the mechanistic steps of nanopore sensing; 2. We will go beyond the current speed and sensitivity limitations of optical single channel recording and develop new single-molecule microscopy methods for nanopore sensing.

基因组测序方法使我们对基因图谱如何影响疾病的微妙和变化的理解迅速扩展。这在一定程度上是受到新技术进步的推动。纳米孔测序已被强调为一种即将到来的技术，有可能走得更远，并提高测序的速度和成本效益。在纳米孔感测中，通过纳米级蛋白质或固态孔的离子流被分析物的存在破坏。如果该分析物是DNA，则检测到对应于该序列的电流中的阻断。无需扩增或标记即可读取数千个碱基的长片段。我们的研究记录标志着我们对开发能够使用荧光显微镜检测单个分子的方法的长期兴趣。我们已经利用这一点来提高我们对膜蛋白功能的理解，特别是专门的蛋白孔如何工作。最近，我们使用这些方法来证明使用光学而不是电读出的纳米孔中的DNA碱基检测。这些方法有可能提高纳米孔传感的并行化。与牛津纳米孔技术公司合作，我们已经确定了当前纳米孔传感的两个基本挑战，其中光学读出提供了显着的影响：1。我们将结合联合收割机光学单通道记录和单分子荧光成像来理解并优化纳米孔传感的机械步骤; 2.我们将超越目前光学单通道记录的速度和灵敏度限制，并开发用于纳米孔传感的新的单分子显微镜方法。

项目成果

Single-molecule imaging of pore-forming toxin dynamics in droplet interface bilayers.

• DOI: 10.1016/bs.mie.2021.01.035
• 发表时间: 2021
• 期刊: Methods in enzymology
• 作者: Christopher Parperis;M. Wallace