Journal Article

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

Wrynla, Xammy Huu, Chen, Xinyi, Stanley, Sarah A., Walker, John A., Liu, Yanxia, Han, Mengting, Chemparathy, Augustine, Jain, Saket, La Russa, Marie F., Rane, Draven A., Abbott, Timothy R., Barron, Annelise E., Lewis, David B., Zeng, Leiping, Lin, Xueqiu, Spatz, Jordan M., Connolly, Michael D., Pinsky, Benjamin, Zhu, Yanyu, Holden, Kevin, Qi, Lei S., Cochran, Jennifer R., Zepeda, Adrianna E., Rustagi, Arjun, Temperton, Nigel, Kadina, Anastasia P., Blish, Catherine A., Wang, Haifeng

2022

Overview

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. A major challenge in coronavirus vaccination and treatment is to counteract rapid viral evolution and mutations. Here the authors show that CRISPR-Cas13d can be used as a broad-spectrum antiviral to inhibit human coronaviruses, including new SARS-CoV-2 variants, combined with small molecule drugs for an enhanced antiviral effect in human primary cells.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 13/109 / 13/31 / 13/44 / 14/19 / 14/32 / 14/35 / 38/77 / 38/91 / 60 APPLIED LIFE SCIENCES / 631/326/596/4130 / 631/337/4041 / 631/553/552 / 631/61/338/552 / Antiviral agents / Antiviral Agents - pharmacology / Antiviral drugs / Coronaviridae / Coronaviruses / COVID-19 / COVID-19 Drug Treatment / CRISPR / CRISPR-Cas systems / Disease prevention / Drugs / Epithelium / Evolution / Humanities and Social Sciences / Humans / Immunization / Lipids / Liposomes / multidisciplinary / Mutation / Nanoparticles / SARS-CoV-2 / SARS-CoV-2 - genetics / Science / Science (multidisciplinary) / Severe acute respiratory syndrome coronavirus 2 / synthetic biology / Vaccination / Viral diseases