DREAM Sentinels: Selection of aptamers that target viral variants with high specificity

为了对抗传染病，该项目将开发一种多功能平台，该平台在生成新的捕获分子时具有更短的周转时间，该捕获分子能够以高亲和力特异性结合新的病毒变体。研究人员将进一步将捕获的分子转化为检测病毒的传感器，并将其转化为阻断分子以中和病毒。目前的选拔过程耗时耗力。该项目的目标是开发一种简化的过程，可以快速靶向具有高亲和力和特异性的新出现的病毒变体。这项工作将提高我们在病毒传感和治疗方面的知识，建立一个快速反应的生物传感/驱动一体化平台，可以很容易地适应未来的传染病。为了增加影响力，调查人员将与当地K-12学生合作开展几个拓展项目，目的是培训学生开发新工具，并鼓励他们走上科学、技术、工程和数学方面的职业道路。适体传感器和治疗方法的发展受到当前黄金标准SELEX（通过指数富集的配体系统进化）工作流程瓶颈的阻碍，特别是反选择过程，其目的是消除与目标结构相似的候选体。虽然反选择是选择针对特定目标的高度特异性适配体的关键，但这是一个耗时，劳动密集型的过程，经常失败。为了解决这个问题，该项目将SELEX工作流程与一种互补的高通量芯片选择方法相结合，该方法可以充分表征库中每个适体变体针对许多结构相似的病毒靶标的结合亲和力、动力学和特异性。这种全分析方法不仅绕过了执行反选择的需要，而且还允许选择能够区分结构相似的病毒靶标的适体。然后，病毒结合适体将与新型荧光适体设计相结合，以创建新的传感器，该传感器在与目标病毒结合时发光，并可转化为病毒抑制剂，结合并阻断SARS-CoV-2刺突蛋白的受体结合域（RBD），使其无法与人类宿主细胞上的血管紧张素转换酶2 （ACE2）受体相互作用。因此，拟议的研究具有解决新的生物威胁的生物传感和生物驱动组件。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

To combat infectious diseases, this project will develop a versatile platform that has a much shorter turn-around time in generating a new capture molecule that specifically binds a new viral variant with high affinity. The investigators will further convert the captured molecule into a sensor for virus detection, and also turn it into a blocking molecule to neutralize the virus. The current selection process is time-consuming and labor intensive. The goal of this project is to develop a streamlined process that can rapidly target emerging viral variants with high affinity and specificity. The proposed work will advance our knowledge in virus sensing and therapeutics, establishing a quick-responsive biosensing/actuating All-In-One platform that can be easily adapted to address future infectious diseases. To increase impact, the investigators will work with local K-12 students in several outreach programs, with the goal of training the students to develop novel tools and encouraging them into a career path in science, technology, engineering and mathematics.Development of aptamer sensors and therapeutics is hampered by the bottleneck in the workflow of current gold standard SELEX (systematic evolution of ligands by exponential enrichment), especially the counterselection process, which aims to eliminate the candidates that bind structurally similar relatives of the target. While the counterselection is key to selecting highly specific aptamers against a specific target, it is a time-consuming, labor-intensive process that often fails. To address this issue, this project will combine SELEX workflow with a complementary high-throughput chip selection approach that can fully characterize the binding affinity, kinetics and specificity of each aptamer variant in the library against a number of structurally similar viral targets. This total-analysis approach not only bypasses the need to perform counterselections but also allows selection of aptamers that can differentiate structurally similar viral targets. The virus-binding aptamers will then be integrated with novel fluorogenic aptamer design to create new sensors that light up upon binding the target viruses and can be turned into virus inhibitors that bind and block the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein from interacting with the angiotensin-converting enzyme 2 (ACE2) receptors on human host cells. The proposed research thus has both biosensing and bioactuation components that address new biological threats.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.