Identification of host cell components essential for the SARS-CoV-2 life cycle

点击翻译按钮获取中文摘要

All viruses are dependent on host cell machinery for aspects of their lifecycle. We plan to identify non-essential host (human) cell components that are required for the life cycle of the SARS-CoV-2 virus. If we are able to identify them, these will provide critical intervention points for developing drugs that could prevent virus infection or hamper/stop viral replication.We will used a genetic approach, mutating each of the 20,000 genes in the genome and testing whether the virus is able to infect the cell, replicate and produce new infectious particles. The results will provide insights into molecular host-pathogen interactions at various steps of the virus life cycle including entrance, replication, biosynthesis, and release. Experimentally, we will try to identify mutant cells which do not support aspects of the viral life cycle. We will then seek to identify the underlying mutant gene(s) which are able provide this protection. Technically this will be achieved by killing/removing cells which have been successfully infected, allowing the "resistant" ones to survive. The initial stage of the project will be to establish sensitive and specific assays in human cell lines which can be successfully infected with SARS-CoV-2. Genetic screens would then be conducted with CRISPR libraries that target all the gene in the genome as well as a library specialised on "drug targetable" host genes. If hits are identified in a "druggable" library, the results could be rapidly translated. For instance by taking the corresponding drug and testing this on cells in culture. Hits identified by genome wide screens will take longer to translate, depending on the gene that is identified and prior work on the gene. The reporter cell lines we are developing to support this project as well as any targets we identify should support collaborative efforts with other laboratories and industrial partners, to translate these findings into host-directed therapies.