Journal Article

A Dual-Reporter System for Real-Time Monitoring and High-throughput CRISPR/Cas9 Library Screening of the Hepatitis C Virus

Cai, Changzu, Zhang, Linqi, Wei, Wensheng, Yuan, Pengfei, Ren, Qingpeng, Li, Chan, Luo, Guangxiang George

2015

Overview

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 ( N S3-4A I nducible r tTA-mediated D ual-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.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 14/10 / 38/43 / 38/47 / 38/70 / 45/77 / 49/31 / 631/61/191/1908 / 692/420/254 / CD81 antigen / Cirrhosis / Clustered Regularly Interspaced Short Palindromic Repeats - genetics / CRISPR / Gene Library / Genes, Reporter - genetics / HeLa Cells / Hepacivirus / Hepatitis / Hepatitis C / Hepatocellular carcinoma / High-Throughput Nucleotide Sequencing - methods / Humanities and Social Sciences / Humans / Liver / Liver cancer / Liver cirrhosis / multidisciplinary / Science / Spectrometry, Fluorescence - methods / Viral Proteins - genetics / Virus Internalization