The production and application of SARS-CoV-2 reverse genetic systems to facilitate vaccine development and biosafe drug discovery platform
- Funded by Department of Health and Social Care / National Institute for Health and Care Research (DHSC-NIHR), UK Research and Innovation (UKRI)
- Total publications:19 publications
Grant number: MR/V027506/1
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Key facts
Disease
COVID-19Start & end year
20202021Known Financial Commitments (USD)
$134,915.59Funder
Department of Health and Social Care / National Institute for Health and Care Research (DHSC-NIHR), UK Research and Innovation (UKRI)Principal Investigator
Dr. Andrew DavidsonResearch Location
United KingdomLead Research Institution
University of BristolResearch Priority Alignment
N/A
Research Category
Therapeutics research, development and implementation
Research Subcategory
Pre-clinical studies
Special Interest Tags
N/A
Study Type
Non-Clinical
Clinical Trial Details
N/A
Broad Policy Alignment
Pending
Age Group
Not Applicable
Vulnerable Population
Not applicable
Occupations of Interest
Not applicable
Abstract
The development of coronavirus reverse genetic systems has been slow and limited to few laboratories worldwide. We have established a rapid approach to generate a SARSCoV-2 reverse genetic system, by targeted recombination of synthetic SARS-CoV-2 cDNA fragments in yeast, using a yeast artificial chromosome (YAC) vector. Nanopore minION sequencing was used to quickly identify correctly assembled YAC clones, that can be transcribed in vitro to produce RNA corresponding to the SARS-CoV-2 genome, which can be used to recover recombinant SARS-CoV-2 (Figure 1A). The entire process takes 2-3 weeks from the receipt of synthetic DNA. We now propose to use this reverse genetic system to develop: 1) recombinant SARS-CoV-2 reporter viruses expressing one or more reporter genes, in place of genes that are non-essential in cell culture: SARS-CoV-2 reporter viruses can be used to rapidly characterise viral infectivity and will be used to develop high-throughput virus antibody neutralisation and escape assays, to underpin viral vaccine development. 2) a biosafe SARS-CoV-2 "replicon" which lacks the viral structural genes but can replicate intracellularly. Human cell lines stably expressing SARS-CoV-2 replicons are powerful tools for antiviral screening and in contrast to assays with SARS-CoV-2, can be undertaken at containment level 2. These constructs will allow academic and commercial laboratories to undertake highthroughput antiviral assays, removing a bottleneck in drug discovery efforts. Furthermore, combined transcriptomics and proteomics will be used to characterise the effects of the recombinant reporter viruses and replicons on the host cell, ensuring that the engineered viruses faithfully represent the natural virus.
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