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

为了对抗传染病，该项目将开发一个多功能平台，在产生新的捕获分子方面具有更短的周转时间，该分子专门结合具有高亲和力的新病毒变体。研究人员将进一步将捕获的分子转化为病毒检测的传感器，并将其转化为阻断分子，以中和病毒。目前的遴选过程既耗时又费力。该项目的目标是开发一种简化的过程，能够快速针对具有高亲和力和特异性的新出现的病毒变体。拟议的工作将促进我们在病毒传感和治疗方面的知识，建立一个快速响应的生物传感/致动一体化平台，可以很容易地适应未来的传染病。为了增加影响力，调查人员将在几个外展项目中与当地的K-12学生合作，目的是培训学生开发新的工具，并鼓励他们进入科学、技术、工程和数学领域的职业道路。适配子传感器和治疗学的开发受到当前黄金标准SELEX(通过指数浓缩的配体的系统进化)工作流程中的瓶颈，特别是反选择过程，该过程旨在消除绑定目标结构相似亲属的候选人。虽然反选择是针对特定靶标选择高度特异性适配子的关键，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工作流程与互补的高通量芯片选择方法相结合，该方法可以充分表征文库中每个适配子变体与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省去了进行反选择的需要，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，病毒结合适配子将与新型荧光适配子设计相结合，以创建新的传感器，该传感器在与目标病毒结合时会发光，并可转变为病毒抑制剂，结合并阻止SARS-CoV-2刺突蛋白的受体结合域(RBD)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，这项拟议的研究同时包含了生物传感和生物驱动组件，以应对新的生物威胁。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.