Pan-Coronavirus CRISPR-CasRx Effector System Significantly Reduces Viable Titer in HCoV-OC43, HCoV-229E, and SARS-CoV-2

Mayes, Cathryn M.; Santarpia, Joshua L.

doi:10.1089/crispr.2022.0095

Pan-Coronavirus CRISPR-CasRx Effector System Significantly Reduces Viable Titer in HCoV-OC43, HCoV-229E, and SARS-CoV-2

Journal Article · Mon Mar 13 00:00:00 EDT 2023 · The CRISPR Journal

DOI:https://doi.org/10.1089/crispr.2022.0095· OSTI ID:2469877

^[1]; Santarpia, Joshua L. ^[2]

Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); National Strategic Research Institute, Omaha, NE (United States)
Univ. of Nebraska, Omaha, NE (United States); National Strategic Research Institute, Omaha, NE (United States)

CRISPR-based technology has become widely used as an antiviral strategy, including as a broad-spectrum human coronavirus (HCoV) therapeutic. In this work, we have designed a CRISPR-CasRx effector system with guide RNAs (gRNAs) that are cross-reactive among several HCoV species. We tested the efficacy of this pan-coronavirus effector system by evaluating the reduction in viral viability associated with different CRISPR targets in HCoV-OC43, HCoV-229E, and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We determined that several CRISPR targets significantly reduce viral titer, despite the presence of single nucleotide polymorphisms in the gRNA when compared with a non-targeting, negative control gRNA. CRISPR targets reduced viral titer between 85% and >99% in HCoV-OC43, between 78% and >99% in HCoV-229E, and between 70% and 94% in SARS-CoV-2 when compared with an untreated virus control. These data establish a proof-of-concept for a pan-coronavirus CRISPR effector system that is capable of reducing viable virus in both Risk Group 2 and Risk Group 3 HCoV pathogens.

View Accepted Manuscript (DOE)

Research Organization:: Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

Sponsoring Organization:: USDOE National Nuclear Security Administration (NNSA)

Grant/Contract Number:: NA0003525

OSTI ID:: 2469877

Journal Information:: The CRISPR Journal, Journal Name: The CRISPR Journal Journal Issue: 4 Vol. 6; ISSN 2573-1599

Publisher:: Mary Ann Liebert, Inc.Copyright Statement

Country of Publication:: United States

Language:: English

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60 APPLIED LIFE SCIENCES
CRISPR
biotechnology
coronaviruses
diagnosis
genetic engineering
genome editing
medicine
pathogens
regenerative medicine
surgery
viruses

Pan-Coronavirus CRISPR-CasRx Effector System Significantly Reduces Viable Titer in HCoV-OC43, HCoV-229E, and SARS-CoV-2

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