A Dual-reporter system for real-time monitoring and high-throughput CRISPR/Cas9 library screening of the hepatitis C virus

Scientific Reports
Qingpeng RenWensheng Wei

Abstract

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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Citations

Nov 6, 2015·Viruses·Mirjam B ZeiselCatherine Schuster
Dec 1, 2015·Biochemical and Biophysical Research Communications·Yaogang ZhongZheng Li
Mar 4, 2015·The FEBS Journal·Jingyu PengWensheng Wei
Jul 3, 2015·Cell & Bioscience·Sean M RiordanShui Qing Ye
Nov 1, 2016·Expert Opinion on Drug Discovery·Liudi TangJu-Tao Guo
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May 18, 2016·Scientific Reports·Jens-Ola Ekström, Dan Hultmark
Jan 18, 2018·Viruses·Shuliang ChenDeyin Guo
Aug 23, 2018·Journal of Medical Virology·Usman GilaniNaveed Shahzad
Jun 25, 2016·Scientific Reports·Jian-Ping ZhangXiao-Bing Zhang
Jun 3, 2016·The FEBS Journal·Linde A MilesJohn T Poirier
Aug 5, 2020·Proceedings of the National Academy of Sciences of the United States of America·Rebecca K LaneWilliam J Kaiser
Dec 20, 2019·Journal of Virology·Isabel GallegoEsteban Domingo
Dec 29, 2020·Nanomedicine : Nanotechnology, Biology, and Medicine·Neelam YadavJogender Singh Rana
Dec 10, 2021·Hormone Molecular Biology and Clinical Investigation·Dhivya SelvarajAnita Devi

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Methods Mentioned

BETA
FACS
gene-knockout
fluorescence microscopy

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