RAPID: Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)-Prevention: Multiple-Site Binding with Fusing Aptamers to mitigate Coronavirus Disease 2019

SARS-CoV-2导致2019年新型冠状病毒传染病(新冠肺炎)。病毒刺突蛋白与受体蛋白(称为血管紧张素转换酶2，或ACE2)的结合是感染人类宿主细胞的第一步。阻止或抑制这种相互作用可以阻止这种高传染性病毒对人类细胞的入侵。通过这一奖项，生命过程化学项目支持俄亥俄州鲍林格林大学的谭晓红博士的研究，以创造DNA适体来抑制病毒入侵的这一初始步骤。适配子是DNA的短序列，可以折叠成与目标分子相匹配的形状，从而阻止其他分子结合。这些DNA适配子是通过从不同序列的大型混合物中连续选择分子而“设计”的，从而产生少数几个序列，阻止所有其他DNA与靶标结合。谭博士使用这种选择过程来寻找与SARS-CoV-2刺突蛋白紧密结合的DNA适配子。更多识别出的DNA适配子中的两个被连接在一起，以产生比任何单一适配子更好地抑制相互作用的“超级结合”。构建阻止SARS-CoV-2入侵人体细胞的DNA适配子可能会导致抗病毒药物的设计，从而帮助降低导致新冠肺炎大流行的传染性。教育和培训活动将重点放在博士后研究人员学习化学生物学前沿技术的前线研究问题上。该团队正在开发利用远程学习技术向公众、K-12机构和其他研究小组传播他们的科学的方法。该项目的目标是开发与SARS-CoV-2结合区域显著相互作用的聚合物分子(融合DNA适配子)。其目的是有效地阻止病毒与特定的血管紧张素转换酶2受体结合，从而阻止病毒进入人类细胞，从而阻止其导致新冠肺炎感染。谭教授使用SELEX(指数富集法配体的系统进化)来识别与SARS-CoV-2尖峰蛋白(S)细胞结合区域上三个不同区域相互作用的单个适配子。适配子与其靶标的亲和力是通过标准结合分析来测量的。这些单独的高亲和力DNA适配子使用核酸或化学连接物连接在一起，形成具有与抗体相似亲和力的融合适配子。使用标准技术与重组SARS-CoV-2刺突蛋白以及与托莱多大学医学院合作的活SARS-CoV-2进行了测试，验证了这些融合DNA适体“超级结合”有效阻止这种相互作用的能力。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。

The SARS-CoV-2 causes the novel coronavirus infectious disease 2019 (COVID-19). The binding of the viral spike protein to a receptor protein (called the angiotensin converting enzyme 2, or ACE2) is the first step in the infection of the human host cell. Blocking or inhibiting this interaction could stop the invasion of the human cell by this highly infectious virus. With this award, the Chemistry of Life Processes program is supporting the research of Dr. Xiaohong Tan at Bowling Green University in Bowling Green, Ohio to create DNA aptamers to inhibit this initial step of viral invasion. Aptamers are short sequences of DNA that fold into a shape that matches that of a target molecule, thereby keeping other molecules from binding. These DNA aptamers are “designed” by successive selection of molecules from large mixtures of different sequences, resulting in a few sequences that keep all other DNAs from binding the target. Dr. Tan uses this selection process to find DNA aptamers that bind tightly to the SARS-CoV-2 spike protein. Two of more of the identified DNA aptamers are linked together to create “super binders” that are much better at inhibiting the interaction than would any single aptamer. Constructing DNA aptamers that block the invasion of SARS-CoV-2 into human cells may lead to the design of antiviral agents that help lessen the infectivity that causes the COVID-19 pandemic. Educational and training activities will focus on postdoctoral researchers learning cutting edge techniques in chemical biology on a “front line” research problem. The team is developing ways to communicate their science to the public, to K-12 institutions, and to other research groups using distance learning technologies.The goal of this project is to develop polymeric molecules (fusion DNA aptamers) that significantly interact with the binding domain of SARS-CoV-2. The objective is to effectively block the virus from binding to the specific ACE2 receptor, thereby blocking the entry of the virus into human cells and, as a result, its ability to cause COVID-19 infections. Professor Tan uses SELEX (systematic evolution of ligands by exponential enrichment) to identify individual aptamers that interact with three distinct regions on the cell binding domain of the spike (S) protein of SARS-CoV-2. The affinities of aptamers to their targets are measured through standard binding assays. These individual high affinity DNA aptamers are connected together using nucleic acid or chemical linkers to make fusion aptamers that have affinities comparable to those of antibodies. The ability of these fused DNA aptamer “super binders” to effectively block this interaction is tested against a recombinant SARS-CoV-2 spike protein using standard technologies, as well as live SARS-CoV-2 through collaboration with the University of Toledo Medical School.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.