Programmable Inhibition and Detection of RNA Viruses Using Cas13.

TitleProgrammable Inhibition and Detection of RNA Viruses Using Cas13.
Publication TypeJournal Article
Year of Publication2019
AuthorsFreije, CA, Myhrvold, C, Boehm, CK, Lin, AE, Welch, NL, Carter, A, Metsky, HC, Luo, CY, Abudayyeh, OO, Gootenberg, JS, Yozwiak, NL, Zhang, F, Sabeti, PC
JournalMol Cell
Date Published2019 Dec 05
KeywordsA549 Cells, Animals, Chlorocebus aethiops, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Associated Proteins, CRISPR-Cas Systems, Dogs, Escherichia coli, Gene Targeting, HEK293 Cells, Humans, Madin Darby Canine Kidney Cells, RNA Stability, RNA Viruses, RNA, Viral, Vero Cells

<p>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.</p>

Alternate JournalMol Cell
PubMed ID31607545
PubMed Central IDPMC7422627
Grant ListR01 MH110049 / MH / NIMH NIH HHS / United States
DP1 HL141201 / HL / NHLBI NIH HHS / United States
R01 HG009761 / HG / NHGRI NIH HHS / United States
U19 AI110818 / AI / NIAID NIH HHS / United States
RM1 HG006193 / HG / NHGRI NIH HHS / United States