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Article

A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry

1
Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095, USA
2
Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA
3
Human Oncology & Pathogenesis Program, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA
4
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA 90095, USA
5
California NanoSystems Institute, University of California, Los Angeles, CA 90095, USA
6
Department of Bioengineering, Samueli School of Engineering, University of California, Los Angeles, CA 90095, USA
7
Jonsson Comprehensive Cancer Center, University of California, Los Angeles, CA 90095, USA
8
Department of Biological Science, Florida State University, Tallahassee, FL 32306, USA
9
Division of Hematology and Oncology, Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA
10
AIDS Institute, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA
11
Druckenmiller Center for Lung Cancer Research and Department of Medicine, Thoracic Oncology Service, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA
12
Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, CA 90095, USA
13
Laura and Isaac Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016, USA
14
Molecular Biology Institute, University of California, Los Angeles, CA 90095, USA
*
Author to whom correspondence should be addressed.
These authors have contributed equally to the study.
Current address: Regeneron Pharmaceuticals, Inc., Tarrytown, NY 10591, USA.
Academic Editors: Jiri Hejnar and Ben Berkhout
Viruses 2021, 13(11), 2113; https://doi.org/10.3390/v13112113
Received: 29 August 2021 / Revised: 12 October 2021 / Accepted: 13 October 2021 / Published: 20 October 2021
(This article belongs to the Special Issue CRISPR/Cas in Viral Research)
Zika virus (ZIKV) is a re-emerging flavivirus that has caused large-scale epidemics. Infection during pregnancy can lead to neurologic developmental abnormalities in children. There is no approved vaccine or therapy for ZIKV. To uncover cellular pathways required for ZIKV that can be therapeutically targeted, we transcriptionally upregulated all known human coding genes with an engineered CRISPR–Cas9 activation complex in human fibroblasts deficient in interferon (IFN) signaling. We identified Ras homolog family member V (RhoV) and WW domain-containing transcription regulator 1 (WWTR1) as proviral factors, and found them to play important roles during early ZIKV infection in A549 cells. We then focused on RhoV, a Rho GTPase with atypical terminal sequences and membrane association, and validated its proviral effects on ZIKV infection and virion production in SNB-19 cells. We found that RhoV promotes infection of some flaviviruses and acts at the step of viral entry. Furthermore, RhoV proviral effects depend on the complete GTPase cycle. By depleting Rho GTPases and related proteins, we identified RhoB and Pak1 as additional proviral factors. Taken together, these results highlight the positive role of RhoV in ZIKV infection and confirm CRISPR activation as a relevant method to identify novel host–pathogen interactions. View Full-Text
Keywords: CRISPR activation; Zika virus; proviral factors; WWTR1; Rho GTPases; RhoV CRISPR activation; Zika virus; proviral factors; WWTR1; Rho GTPases; RhoV
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MDPI and ACS Style

Luu, A.P.; Yao, Z.; Ramachandran, S.; Azzopardi, S.A.; Miles, L.A.; Schneider, W.M.; Hoffmann, H.-H.; Bozzacco, L.; Garcia, G., Jr.; Gong, D.; Damoiseaux, R.; Tang, H.; Morizono, K.; Rudin, C.M.; Sun, R.; Arumugaswami, V.; Poirier, J.T.; MacDonald, M.R.; Rice, C.M.; Li, M.M.H. A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry. Viruses 2021, 13, 2113. https://doi.org/10.3390/v13112113

AMA Style

Luu AP, Yao Z, Ramachandran S, Azzopardi SA, Miles LA, Schneider WM, Hoffmann H-H, Bozzacco L, Garcia G Jr., Gong D, Damoiseaux R, Tang H, Morizono K, Rudin CM, Sun R, Arumugaswami V, Poirier JT, MacDonald MR, Rice CM, Li MMH. A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry. Viruses. 2021; 13(11):2113. https://doi.org/10.3390/v13112113

Chicago/Turabian Style

Luu, Anh Phuong, Zhenlan Yao, Sangeetha Ramachandran, Stephanie A. Azzopardi, Linde A. Miles, William M. Schneider, H.-Heinrich Hoffmann, Leonia Bozzacco, Gustavo Garcia Jr., Danyang Gong, Robert Damoiseaux, Hengli Tang, Kouki Morizono, Charles M. Rudin, Ren Sun, Vaithilingaraja Arumugaswami, John T. Poirier, Margaret R. MacDonald, Charles M. Rice, and Melody M. H. Li. 2021. "A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry" Viruses 13, no. 11: 2113. https://doi.org/10.3390/v13112113

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