Global genome screening with CRISPRko libraries to identify essential factors in SARS-CoV2 infection and replication

Abstract

This decade begins with a worldwide health emergency, the new Coronavirus pandemic, cause of Severe Acute Respiratory Syndrome 2 (SARS-CoV2). Until this date, there are more than 900,000 positive cases that have resulted in more than 44,000 deaths in 180 countries. In view of these epidemiological data, strategies are needed to contain the transmission of the virus and mainly effective alternatives in the treatment of this disease. In this context, with this project we aim to evaluate the versatility of CRISPRko library in the identification of critical factors for SARS-Cov2 infection and replication. This library consists of a group of 77,441 guides directed to approximately 19 thousand human genes. This tool will allow us to globally screen the human genome, through gene deletion mediated by CRISPR/Cas9 in human lung epithelial cells. After selection against essentialgenes for cell viability, we will submit the cells to five rounds of lethal infection with human isolate strain (from Brazil) of SARS-Cov2. Finally, we will be able to determine genes and pathways involved in viral replication and infection, by sequencing the surviving cells and detecting the guides present in that population. With this high coverage strategy, we believe that we will be able to point out new potential targets for the development of therapies for this disease. (AU)