Programmable Inhibition and Detection of RNA Viruses Using Cas13

Overview

Journal Mol Cell

Publisher Cell Press

Specialty Cell Biology

Date 2019 Oct 15

PMID 31607545

Citations 188

Authors

Catherine A Freije

Cameron Myhrvold

Chloe K Boehm

Aaron E Lin

Nicole L Welch

Amber Carter

Hayden C Metsky

Cynthia Y Luo

Omar O Abudayyeh

Jonathan S Gootenberg

Nathan L Yozwiak

Feng Zhang

Pardis C Sabeti

Affiliations

Soon will be listed here.

Abstract

The CRISPR effector Cas13 could be an effective antiviral for single-stranded RNA (ssRNA) viruses because it programmably cleaves RNAs complementary to its CRISPR RNA (crRNA). Here, we computationally identify thousands of potential Cas13 crRNA target sites in hundreds of ssRNA viral species that can potentially infect humans. We experimentally demonstrate Cas13's potent activity against three distinct ssRNA viruses: lymphocytic choriomeningitis virus (LCMV); influenza A virus (IAV); and vesicular stomatitis virus (VSV). Combining this antiviral activity with Cas13-based diagnostics, we develop Cas13-assisted restriction of viral expression and readout (CARVER), an end-to-end platform that uses Cas13 to detect and destroy viral RNA. We further screen hundreds of crRNAs along the LCMV genome to evaluate how conservation and target RNA nucleotide content influence Cas13's antiviral activity. Our results demonstrate that Cas13 can be harnessed to target a wide range of ssRNA viruses and CARVER's potential broad utility for rapid diagnostic and antiviral drug development.

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