Key result
Using the novel NIrD dual-reporter system and a focused CRISPR/Cas9 library, CLDN1, OCLN, and CD81 were identified as essential genes for both cell-free entry and cell-to-cell transmission of HCV.
The NIrD dual-reporter system combined with CRISPR/Cas9 screening successfully identifies critical host factors for HCV infection, such as CLDN1, OCLN, and CD81.
The hepatitis C virus (HCV) is one of the leading causes of chronic hepatitis, liver cirrhosis and hepatocellular carcinomas and infects approximately 170 million people worldwide. Although several reporter systems have been developed, many shortcomings limit their use in the assessment of HCV infections. Here, we report a real-time live-cell reporter, termed the NIrD (NS3-4A Inducible rtTA-mediated Dual-reporter) system, which provides an on-off switch specifically in response to an HCV infection. Using the NIrD system and a focused CRISPR/Cas9 library, we identified CLDN1, OCLN and CD81 as essential genes for both the cell-free entry and the cell-to-cell transmission of HCV. The combination of this ultra-sensitive reporter system and the CRISPR knockout screening provides a powerful and high-throughput strategy for the identification of critical host components for HCV infections.
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Ren et al. (2015) studied Hepatitis C Virus (HCV) infection. NIrD (NS3-4A Inducible rtTA-mediated Dual-reporter) system and CRISPR/Cas9 screening was evaluated on Identification of essential host genes for HCV infection. Using the novel NIrD dual-reporter system and a focused CRISPR/Cas9 library, CLDN1, OCLN, and CD81 were identified as essential genes for both cell-free entry and cell-to-cell transmission of HCV.
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