The production and application of SARS-CoV-2 reverse genetic systems to facilitate vaccine development and biosafe drug discovery platforms.

SARS-CoV-2反向遗传系统的生产和应用，以促进疫苗开发和生物安全药物发现平台。

• 批准号: MR/V027506/1
• 负责人: Andrew Davidson
• 金额: $ 26.25万
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FV027506%2F1
• 关键词: production; application; SARS; CoV; reverse

The development of vaccines and antiviral drugs against SARS-CoV-2 is hampered by the requirement to grow the virus under high-containment, which is limited to few laboratories worldwide. We have recently established a rapid procedure to produce a DNA copy of the SARS-CoV-2 RNA genome, that can be used to genetically engineer new viruses via a process termed "reverse genetics". We now propose to engineer the SARS-CoV-2 genome to produce 1) a SARS-CoV-2 virus that expresses a green fluorescent protein (turboGFP) and 2) a disabled biosafe form of the virus (replicon) that expresses turboGFP instead of the structurally important spike protein. When introduced into suitable cells, the replicon reproduces continously but does not produce infectious virus particles. The two engineered forms of the virus can then be used to develop high-throughput fluorescence based assays to monitor the entry of the virus into cells and its replication. The first can be used to identify antibodies induced by vaccine candidates that neutralise virus infectivity, and also viral mutations that allow escape from the antibody response. The replicon can be used for rapid screening of antiviral compounds, at a lower containment level, which would allow many more laboratories to participate in drug discovery efforts.

针对SARS-CoV-2的疫苗和抗病毒药物的开发受到限制，因为需要在高度封闭的条件下培养病毒，而这仅限于全世界少数实验室。我们最近建立了一个快速的程序来生产SARS-CoV-2 RNA基因组的DNA拷贝，可以通过称为“反向遗传学”的过程来遗传工程新病毒。我们现在建议对SARS-CoV-2基因组进行工程改造，以产生1）表达绿色荧光蛋白（turboGFP）的SARS-CoV-2病毒和2）表达turboGFP而不是结构上重要的刺突蛋白的病毒的禁用生物安全形式（复制子）。当引入合适的细胞中时，复制子连续复制，但不产生感染性病毒颗粒。然后，可以使用病毒的两种工程形式来开发基于荧光的高通量测定，以监测病毒进入细胞及其复制。第一种方法可用于鉴定候选疫苗诱导的抗体，这些抗体可中和病毒感染性，也可用于鉴定病毒突变，这些突变可使病毒逃避抗体反应。复制子可用于快速筛选抗病毒化合物，在较低的遏制水平，这将允许更多的实验室参与药物发现的努力。

项目成果

Recombinant SARS-CoV-2 lacking initiating and internal methionine codons within ORF10 is attenuated in vivo

• DOI: 10.1101/2023.08.04.551973
• 发表时间: 2023-08
• 期刊: bioRxiv
• 作者: Shichun Gu;Eleanor G. Bentley;R. Milligan;Abdulaziz M Almuqrin;Parul Sharma;Adam Kirby;D. F. Mega;A. Kipar;Maximilian Erdmann;J.-M. Bazire;K. Heesom;Philip A. Lewis;I. Donovan-Banfield;Charlotte Reston;Isobel Webb;Simon de Neck;Xaiofeng Dong;J. Hiscox;A. Davidson;James P. Stewart;D. Matthews

In vitro generated antibodies guide thermostable ADDomer nanoparticle design for nasal vaccination and passive immunization against SARS-CoV-2

• DOI: 10.1093/abt/tbad024
• 发表时间: 2023-11-10
• 期刊: ANTIBODY THERAPEUTICS
• 作者: Buzas, Dora;Bunzel, Adrian H.;Berger, Imre
• 通讯作者: Berger, Imre

Neuropilin-1 is a host factor for SARS-CoV-2 infection.

• DOI: 10.1126/science.abd3072
• 发表时间: 2020-11-13
• 期刊: Science (New York, N.Y.)
• 作者: Daly JL;Simonetti B;Klein K;Chen KE;Williamson MK;Antón-Plágaro C;Shoemark DK;Simón-Gracia L;Bauer M;Hollandi R;Greber UF;Horvath P;Sessions RB;Helenius A;Hiscox JA;Teesalu T;Matthews DA;Davidson AD;Collins BM;Cullen PJ;Yamauchi Y
• 通讯作者: Yamauchi Y

Post-acute COVID-19 associated with evidence of bystander T-cell activation and a recurring antibiotic-resistant bacterial pneumonia.

• DOI: 10.7554/elife.63430
• 发表时间: 2020-12-17
• 期刊: eLife
• 影响因子: 7.7
• 作者: Gregorova M;Morse D;Brignoli T;Steventon J;Hamilton F;Albur M;Arnold D;Thomas M;Halliday A;Baum H;Rice C;Avison MB;Davidson AD;Santopaolo M;Oliver E;Goenka A;Finn A;Wooldridge L;Amulic B;Boyton RJ;Altmann DM;Butler DK;McMurray C;Stockton J;Nicholls S;Cooper C;Loman N;Cox MJ;Rivino L;Massey RC
• 通讯作者: Massey RC

The P323L substitution in the SARS-CoV-2 polymerase (NSP12) confers a selective advantage during infection.

• DOI: 10.1186/s13059-023-02881-5
• 发表时间: 2023-03-13
• 期刊: Genome biology
• 影响因子: 12.3