@article{ec58d3857d27427991ffd9c0c42fd63e,
title = "Inducible CRISPR activation screen for interferon-stimulated genes identifies OAS1 as a SARS-CoV-2 restriction factor",
abstract = "Interferons establish an antiviral state through the induction of hundreds of interferon-stimulated genes (ISGs). The mechanisms and viral specificities for most ISGs remain incompletely understood. To enable high-throughput interrogation of ISG antiviral functions in pooled genetic screens while mitigating potentially confounding effects of endogenous interferon and antiproliferative/proapoptotic ISG activities, we adapted a CRISPR-activation (CRISPRa) system for inducible ISG expression in isogenic cell lines with and without the capacity to respond to interferons. We used this platform to screen for ISGs that restrict SARS-CoV-2. Results included ISGs previously described to restrict SARS-CoV-2 and novel candidate antiviral factors. We validated a subset of these by complementary CRISPRa and cDNA expression experiments. OAS1, a top-ranked hit across multiple screens, exhibited strong antiviral effects against SARS-CoV-2, which required OAS1 catalytic activity. These studies demonstrate a high-throughput approach to assess antiviral functions within the ISG repertoire, exemplified by identification of multiple SARS-CoV-2 restriction factors.",
author = "Oded Danziger and Patel, {Roosheel S.} and DeGrace, {Emma J.} and Rosen, {Mikaela R.} and Rosenberg, {Brad R.}",
note = "Funding Information: This work was supported in part by National Institutes of Health grants R01 AI151029 (B.R.R.) and U01 AI150748 (B.R.R.), and funds from the Icahn School of Medicine at Mount Sinai (B.R.R.). Research reported in this paper was also supported by the Office of Research Infrastructure of the National Institutes of Health under award numbers S10OD018522 and S10OD026880. The content is solely the responsibility of the authors and does not necessarily represent the official views of the funders, including the National Institutes of Health. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank Mayte Suarez-Farinas for advice on statistics and experimental design, and Thomas Moran, Center for Therapeutic Antibody Discovery at the Icahn School of Medicine at Mount Sinai, for kindly providing anti-SARS-CoV Nucleocapsid antibody. We thank Meike Dittmann for A549ΔSTAT1 cell lines, Benjamin R. tenOever and Skyler Uhl for technical assistance with plaque assays, and Michael A. Schotsaert for flow cytometry support. We also thank Randy Albrecht for BSL3 facility management and support. We thank Dusan Bogunovic and Ivan Marazzi for discussion, helpful advice, and critical reading of the manuscript. We thank all members of the Rosenberg lab for advice and support. Figure panels Figs 1A and 2A were created with BioRender.com. Publisher Copyright: Copyright: {\textcopyright} 2022 Danziger et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",
year = "2022",
month = apr,
doi = "10.1371/journal.ppat.1010464",
language = "English",
volume = "18",
journal = "PLoS Pathogens",
issn = "1553-7366",
publisher = "Public Library of Science",
number = "4",
}