Programmable Inhibition and Detection of RNA Viruses Using Cas13
© 2019 The Authors 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 vi...
| Main Authors: | , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
Elsevier BV
2021
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| Online Access: | https://hdl.handle.net/1721.1/138385 |
| _version_ | 1826215894889529344 |
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| author | Freije, Catherine A Myhrvold, Cameron Boehm, Chloe K Lin, Aaron E Welch, Nicole L Carter, Amber Metsky, Hayden C Luo, Cynthia Y Abudayyeh, Omar O Gootenberg, Jonathan S Yozwiak, Nathan L Zhang, Feng Sabeti, Pardis C |
| author2 | Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science |
| author_facet | Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Freije, Catherine A Myhrvold, Cameron Boehm, Chloe K Lin, Aaron E Welch, Nicole L Carter, Amber Metsky, Hayden C Luo, Cynthia Y Abudayyeh, Omar O Gootenberg, Jonathan S Yozwiak, Nathan L Zhang, Feng Sabeti, Pardis C |
| author_sort | Freije, Catherine A |
| collection | MIT |
| description | © 2019 The Authors 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. Freije et al. demonstrate that Cas13 can be programmed to target and destroy the genomes of diverse mammalian single-stranded RNA viruses. They identify design principles for efficient Cas13 targeting of viral RNA and create companion Cas13-based diagnostics to rapidly measure the effects of Cas13 targeting. |
| first_indexed | 2024-09-23T16:38:44Z |
| format | Article |
| id | mit-1721.1/138385 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T16:38:44Z |
| publishDate | 2021 |
| publisher | Elsevier BV |
| record_format | dspace |
| spelling | mit-1721.1/1383852023-12-08T19:49:36Z Programmable Inhibition and Detection of RNA Viruses Using Cas13 Freije, Catherine A Myhrvold, Cameron Boehm, Chloe K Lin, Aaron E Welch, Nicole L Carter, Amber Metsky, Hayden C Luo, Cynthia Y Abudayyeh, Omar O Gootenberg, Jonathan S Yozwiak, Nathan L Zhang, Feng Sabeti, Pardis C Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science McGovern Institute for Brain Research at MIT Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences Massachusetts Institute of Technology. Department of Biological Engineering Harvard University--MIT Division of Health Sciences and Technology © 2019 The Authors 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. Freije et al. demonstrate that Cas13 can be programmed to target and destroy the genomes of diverse mammalian single-stranded RNA viruses. They identify design principles for efficient Cas13 targeting of viral RNA and create companion Cas13-based diagnostics to rapidly measure the effects of Cas13 targeting. 2021-12-08T17:59:51Z 2021-12-08T17:59:51Z 2019 2021-12-08T17:51:49Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/138385 Freije, Catherine A, Myhrvold, Cameron, Boehm, Chloe K, Lin, Aaron E, Welch, Nicole L et al. 2019. "Programmable Inhibition and Detection of RNA Viruses Using Cas13." Molecular Cell, 76 (5). en 10.1016/J.MOLCEL.2019.09.013 Molecular Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier BV Elsevier |
| spellingShingle | Freije, Catherine A Myhrvold, Cameron Boehm, Chloe K Lin, Aaron E Welch, Nicole L Carter, Amber Metsky, Hayden C Luo, Cynthia Y Abudayyeh, Omar O Gootenberg, Jonathan S Yozwiak, Nathan L Zhang, Feng Sabeti, Pardis C Programmable Inhibition and Detection of RNA Viruses Using Cas13 |
| title | Programmable Inhibition and Detection of RNA Viruses Using Cas13 |
| title_full | Programmable Inhibition and Detection of RNA Viruses Using Cas13 |
| title_fullStr | Programmable Inhibition and Detection of RNA Viruses Using Cas13 |
| title_full_unstemmed | Programmable Inhibition and Detection of RNA Viruses Using Cas13 |
| title_short | Programmable Inhibition and Detection of RNA Viruses Using Cas13 |
| title_sort | programmable inhibition and detection of rna viruses using cas13 |
| url | https://hdl.handle.net/1721.1/138385 |
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