Programmable Inhibition and Detection of RNA Viruses Using Cas13. Author Catherine Freije, Cameron Myhrvold, Chloe Boehm, Aaron Lin, Nicole Welch, Amber Carter, Hayden Metsky, Cynthia Luo, Omar Abudayyeh, Jonathan Gootenberg, Nathan Yozwiak, Feng Zhang, Pardis Sabeti Publication Year 2019 Type Journal Article 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. Keywords Animals, Escherichia coli, RNA, Viral, Humans, HEK293 Cells, RNA Stability, Vero Cells, Dogs, CRISPR-Cas Systems, Clustered Regularly Interspaced Short Palindromic Repeats, Gene Targeting, Madin Darby Canine Kidney Cells, CRISPR-Associated Proteins, A549 Cells, Chlorocebus aethiops, RNA Viruses Journal Mol Cell Volume 76 Issue 5 Pages 826-837.e11 Date Published 2019 Dec 05 ISSN Number 1097-4164 DOI 10.1016/j.molcel.2019.09.013 Alternate Journal Mol Cell PMCID PMC7422627 PMID 31607545 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML