Broad-spectrum CRISPR-mediated inhibition of SARS-CoV-2 variants and endemic coronaviruses in vitro

Abstract A major challenge in coronavirus vaccination and treatment is to counteract rapid viral evolution and mutations. Here we demonstrate that CRISPR-Cas13d offers a broad-spectrum antiviral (BSA) to inhibit many SARS-CoV-2 variants and diverse human coronavirus strains with >99% reduction of...

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Main Authors: Leiping Zeng, Yanxia Liu, Xammy Huu Wrynla, Timothy R. Abbott, Mengting Han, Yanyu Zhu, Augustine Chemparathy, Xueqiu Lin, Xinyi Chen, Haifeng Wang, Draven A. Rane, Jordan M. Spatz, Saket Jain, Arjun Rustagi, Benjamin Pinsky, Adrianna E. Zepeda, Anastasia P. Kadina, John A. Walker, Kevin Holden, Nigel Temperton, Jennifer R. Cochran, Annelise E. Barron, Michael D. Connolly, Catherine A. Blish, David B. Lewis, Sarah A. Stanley, Marie F. La Russa, Lei S. Qi
Format: Article
Language:English
Published: Nature Portfolio 2022-05-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-022-30546-7
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author Leiping Zeng
Yanxia Liu
Xammy Huu Wrynla
Timothy R. Abbott
Mengting Han
Yanyu Zhu
Augustine Chemparathy
Xueqiu Lin
Xinyi Chen
Haifeng Wang
Draven A. Rane
Jordan M. Spatz
Saket Jain
Arjun Rustagi
Benjamin Pinsky
Adrianna E. Zepeda
Anastasia P. Kadina
John A. Walker
Kevin Holden
Nigel Temperton
Jennifer R. Cochran
Annelise E. Barron
Michael D. Connolly
Catherine A. Blish
David B. Lewis
Sarah A. Stanley
Marie F. La Russa
Lei S. Qi
author_facet Leiping Zeng
Yanxia Liu
Xammy Huu Wrynla
Timothy R. Abbott
Mengting Han
Yanyu Zhu
Augustine Chemparathy
Xueqiu Lin
Xinyi Chen
Haifeng Wang
Draven A. Rane
Jordan M. Spatz
Saket Jain
Arjun Rustagi
Benjamin Pinsky
Adrianna E. Zepeda
Anastasia P. Kadina
John A. Walker
Kevin Holden
Nigel Temperton
Jennifer R. Cochran
Annelise E. Barron
Michael D. Connolly
Catherine A. Blish
David B. Lewis
Sarah A. Stanley
Marie F. La Russa
Lei S. Qi
author_sort Leiping Zeng
collection DOAJ
description Abstract A major challenge in coronavirus vaccination and treatment is to counteract rapid viral evolution and mutations. Here we demonstrate that CRISPR-Cas13d offers a broad-spectrum antiviral (BSA) to inhibit many SARS-CoV-2 variants and diverse human coronavirus strains with >99% reduction of the viral titer. We show that Cas13d-mediated coronavirus inhibition is dependent on the crRNA cellular spatial colocalization with Cas13d and target viral RNA. Cas13d can significantly enhance the therapeutic effects of diverse small molecule drugs against coronaviruses for prophylaxis or treatment purposes, and the best combination reduced viral titer by over four orders of magnitude. Using lipid nanoparticle-mediated RNA delivery, we demonstrate that the Cas13d system can effectively treat infection from multiple variants of coronavirus, including Omicron SARS-CoV-2, in human primary airway epithelium air-liquid interface (ALI) cultures. Our study establishes CRISPR-Cas13 as a BSA which is highly complementary to existing vaccination and antiviral treatment strategies.
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institution Kabale University
issn 2041-1723
language English
publishDate 2022-05-01
publisher Nature Portfolio
record_format Article
series Nature Communications
spelling doaj-art-b19524c1f85b4105af7741efdca8ff412025-08-20T03:45:32ZengNature PortfolioNature Communications2041-17232022-05-0113111610.1038/s41467-022-30546-7Broad-spectrum CRISPR-mediated inhibition of SARS-CoV-2 variants and endemic coronaviruses in vitroLeiping Zeng0Yanxia Liu1Xammy Huu Wrynla2Timothy R. Abbott3Mengting Han4Yanyu Zhu5Augustine Chemparathy6Xueqiu Lin7Xinyi Chen8Haifeng Wang9Draven A. Rane10Jordan M. Spatz11Saket Jain12Arjun Rustagi13Benjamin Pinsky14Adrianna E. Zepeda15Anastasia P. Kadina16John A. Walker17Kevin Holden18Nigel Temperton19Jennifer R. Cochran20Annelise E. Barron21Michael D. Connolly22Catherine A. Blish23David B. Lewis24Sarah A. Stanley25Marie F. La Russa26Lei S. Qi27Department of Bioengineering, Stanford UniversityDepartment of Bioengineering, Stanford UniversityDepartment of Molecular and Cellular Biology, University of CaliforniaDepartment of Bioengineering, Stanford UniversityDepartment of Bioengineering, Stanford UniversityDepartment of Bioengineering, Stanford UniversityDepartment of Bioengineering, Stanford UniversityDepartment of Bioengineering, Stanford UniversityDepartment of Bioengineering, Stanford UniversityDepartment of Bioengineering, Stanford UniversityDepartment of Bioengineering, Stanford UniversityDepartment of Pediatrics, Stanford UniversityUniversity of California San FranciscoDepartment of Medicine, Stanford UniversityDepartment of Medicine, Stanford UniversitySynthego CorporationSynthego CorporationSynthego CorporationSynthego CorporationViral Pseudotype Unit, Medway School of PharmacyDepartment of Bioengineering, Stanford UniversityDepartment of Bioengineering, Stanford UniversityLawrence Berkeley National LaboratoryDepartment of Medicine, Stanford UniversityDepartment of Pediatrics, Stanford UniversityDepartment of Molecular and Cellular Biology, University of CaliforniaDepartment of Bioengineering, Stanford UniversityDepartment of Bioengineering, Stanford UniversityAbstract A major challenge in coronavirus vaccination and treatment is to counteract rapid viral evolution and mutations. Here we demonstrate that CRISPR-Cas13d offers a broad-spectrum antiviral (BSA) to inhibit many SARS-CoV-2 variants and diverse human coronavirus strains with >99% reduction of the viral titer. We show that Cas13d-mediated coronavirus inhibition is dependent on the crRNA cellular spatial colocalization with Cas13d and target viral RNA. Cas13d can significantly enhance the therapeutic effects of diverse small molecule drugs against coronaviruses for prophylaxis or treatment purposes, and the best combination reduced viral titer by over four orders of magnitude. Using lipid nanoparticle-mediated RNA delivery, we demonstrate that the Cas13d system can effectively treat infection from multiple variants of coronavirus, including Omicron SARS-CoV-2, in human primary airway epithelium air-liquid interface (ALI) cultures. Our study establishes CRISPR-Cas13 as a BSA which is highly complementary to existing vaccination and antiviral treatment strategies.https://doi.org/10.1038/s41467-022-30546-7
spellingShingle Leiping Zeng
Yanxia Liu
Xammy Huu Wrynla
Timothy R. Abbott
Mengting Han
Yanyu Zhu
Augustine Chemparathy
Xueqiu Lin
Xinyi Chen
Haifeng Wang
Draven A. Rane
Jordan M. Spatz
Saket Jain
Arjun Rustagi
Benjamin Pinsky
Adrianna E. Zepeda
Anastasia P. Kadina
John A. Walker
Kevin Holden
Nigel Temperton
Jennifer R. Cochran
Annelise E. Barron
Michael D. Connolly
Catherine A. Blish
David B. Lewis
Sarah A. Stanley
Marie F. La Russa
Lei S. Qi
Broad-spectrum CRISPR-mediated inhibition of SARS-CoV-2 variants and endemic coronaviruses in vitro
Nature Communications
title Broad-spectrum CRISPR-mediated inhibition of SARS-CoV-2 variants and endemic coronaviruses in vitro
title_full Broad-spectrum CRISPR-mediated inhibition of SARS-CoV-2 variants and endemic coronaviruses in vitro
title_fullStr Broad-spectrum CRISPR-mediated inhibition of SARS-CoV-2 variants and endemic coronaviruses in vitro
title_full_unstemmed Broad-spectrum CRISPR-mediated inhibition of SARS-CoV-2 variants and endemic coronaviruses in vitro
title_short Broad-spectrum CRISPR-mediated inhibition of SARS-CoV-2 variants and endemic coronaviruses in vitro
title_sort broad spectrum crispr mediated inhibition of sars cov 2 variants and endemic coronaviruses in vitro
url https://doi.org/10.1038/s41467-022-30546-7
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