Development of a virus-free sensor system to repurpose approved drugs for blocking Coronavirus replication

开发无病毒传感器系统，以重新利用已批准的药物来阻止冠状病毒复制

• 批准号: BB/V017780/1
• 负责人: Betty Chung
• 金额: $ 30万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FV017780%2F1
• 关键词: Development; virus; free; sensor; system

The COVID19 pandemic has highlighted the importance of timely discovery of antiviral therapeutics, especially when there is little information as to whether vaccines developed (and pending approval) enable a significant period of immunity. Therefore the development of cost effective, high-throughput sensor systems to rapidly repurpose approved drugs as antiviral agents is extremely desirable. We have developed a virus-free sensor system - pShiftSensor that can specifically screen for drugs to block SARS-CoV2 replication by inhibiting the ribosomal frameshift that is essential for production of the viral replicase. The sensor system is fluorescent-based, therefore permitting cost-effective high-throughput quantitate readout.The system is based on the cis-element required for -1 programmed ribosomal frameshifting (-1 PRF), a non-canonical translation mechanism that is ubiquitous throughout all coronaviruses and used to synthesis viral proteins including the RNA-dependent RNA polymerase (RdRP), essential for viral replication. We have developed pShiftSensor-v1, which is a dual-reporter system that contains the SARS-CoV-2 -1PRF cassette (i.e. the slippery sequence UUUAAAC and the 3' pseudoknot). The system is bi-cistronic thereby avoiding expression artifacts due to differential delivery between the control and test reporter expression cassette. Through this proposal, we will upgrade our system to pShiftSensor-V2, which targets primary cell types such as small airway lung cells, the key entry cell type for SARS-CoV2 infection, and primary leukocytes, especially monocytes/macrophages and T-cells, essential for systemic infection. pShiftSensor-V2 will permit cost-effective, high-throughput cell-type specific screening of viral replication inhibitory compounds in a physiological environment.

COVID 19大流行凸显了及时发现抗病毒疗法的重要性，特别是在关于开发的疫苗（以及正在等待批准的疫苗）是否能够有效免疫的信息很少的情况下。因此，开发具有成本效益的高通量传感器系统以快速地将批准的药物重新用作抗病毒剂是极其期望的。我们已经开发了一种无病毒的传感器系统- pShiftSensor，它可以通过抑制核糖体移码来特异性地筛选阻断SARS-CoV 2复制的药物，核糖体移码是产生病毒复制酶所必需的。该传感器系统基于-1程序化核糖体移码（-1 PRF）所需的顺式元件，这是一种在所有冠状病毒中普遍存在的非经典翻译机制，用于合成病毒蛋白，包括病毒复制所必需的RNA依赖性RNA聚合酶（RdRP）。我们已经开发了pShiftSensor-v1，这是一个双报告系统，包含SARS-CoV-2 - 1 PRF盒（即滑动序列UUUAAAC和3'假结）。该系统是双顺反子的，从而避免了由于对照和测试报告基因表达盒之间的差异递送而导致的表达假象。通过该提案，我们将把我们的系统升级到pShiftSensor-V2，它针对的是原代细胞类型，例如小气道肺细胞（SARS-CoV 2感染的关键进入细胞类型）和原代白细胞，特别是单核细胞/巨噬细胞和T细胞，对于系统性感染至关重要。pShiftSensor-V2将允许在生理环境中对病毒复制抑制化合物进行成本有效的、高通量的细胞类型特异性筛选。

项目成果

RNA structure mediated thermoregulation: What can we learn from plants?

RNA结构介导的温度调节：我们可以从植物中学到什么？

• DOI: 10.17863/cam.88256
• 发表时间: 2022
• 作者: Thomas S

Pervasive translational control of photosynthesis genes during photomorphogenesis is acquired by C 4 genes

光形态发生过程中光合作用基因的普遍翻译控制是由 C 4 基因获得的

• DOI: 10.1101/2023.10.27.563924
• 发表时间: 2023
• 作者: Reyna-Llorens I

The distinct translational landscapes of Gram-positive and Gram-negative bacteria

革兰氏阳性菌和革兰氏阴性菌的独特翻译景观

• DOI: 10.1101/2023.05.25.542305
• 发表时间: 2023
• 作者: Bryant O

The distinct translational landscapes of gram-negative Salmonella and gram-positive Listeria.

• DOI: 10.1038/s41467-023-43759-1
• 发表时间: 2023-12-09
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Bryant, Owain J.;Lastovka, Filip;Powell, Jessica;Chung, Betty Y. -W.
• 通讯作者: Chung, Betty Y. -W.