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“Genetic scissors” against SARS-CoV-2

“Genetic scissors” against SARS-CoV-2

Researchers from the JU Małopolska Centre of Biotechnology have analysed the SARS-CoV-2 infetion using the CRISPR-Cas9 system, known as “genetic scissors”. The results of the research, published in Cells provide new interesting information about the virus replication. Aleksandra Synowiec from the Virogenetics lab at the JU MCB offers more details on this issue.

During the COVID-19 pandemic, virology has become one of the most rapidly developing scientific disciplines and the results of the research in this area have a direct impact on the  approach to the pandemic. Learning the molecular mechanisms of infection means answering the question how SARS-CoV-2 infects our cells and causes illness, which is the first step towards creating effective drugs and treatment strategies.

The research discussed in the recently published paper used advanced molecular biology and bioinformatic analysis methods to identify new cellular factors required for viral replication. The scientists applied CRISPR/Cas9 knock-out screening method using the CRISPR/Cas9 technology, based on the system of defence against mobile genetic elements in bacteria. In this system, Cas proteins conjoined with short RNA fragments (gRNA; guide RNA) sever DNA sequences located in cells. Transferring this defence system to eucaryotic cells (e.g. human cells) allows effective gene editing. The technology was used to create the CRISPR/Cas9 knock-out library, including gRNA, capable of “switching off” virtually all known genes in human cells. This is a powerful tool allowing the identification of interactions between the virus and cell.

Within the framework of research discussed in the paper, studies were conducted using a HeLa modified cell line, which is a widely used model to analyse SARS-CoV-2 infection. The researchers managed to identify 178 genes. Then, more detailed analyses enabled them to identify 5 factors contributing to the reduction or delay of the virus’ replication: EPGN, USP17, MACF1, PCDHGA1, and GAGE1. An interesting example is provided by MACF1 protein, which takes part in numerous processes and signalling networks related to cell polarization and intracellular transport. The protein is also interesting because of its involvement in Wnt/β-catenin signalling factors, whose regulation is observed during an inflammation and the so-called cytokine storm in COVID-19 patients. Another identified protein, EPGN, is a ligand for the EGFR receptor, which plays a key role in cell differentiation. It has been indicated that EGFR receptor is involved in SARS-CoV-2 replication.

Original text by Aleksandra Synowiec: www.nauka.uj.edu.pl

The paper published in Cells: Synowiec, A.; Jedrysik, M.; Branicki, W.; Klajmon, A.; Lei, J.; Owczarek, K.; Suo, C.; Szczepanski, A.; Wang, J.; Zhang, P.; Labaj, P.P.; Pyrc, K. Identification of Cellular Factors Required for SARS-CoV-2 Replication. Cells 2021, 10, 3159, https://doi.org/10.3390/cells10113159.

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