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Inducing circular RNA formation using the CRISPR endoribonuclease Csy4
Circular RNAs (circRNAs) are highly stable, covalently closed RNAs that are regulated in a spatiotemporal manner and whose functions are largely unknown. These molecules have the potential to be incorporated into engineered systems with broad technological implications. Here we describe a switch for...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5393173/ https://www.ncbi.nlm.nih.gov/pubmed/28223408 http://dx.doi.org/10.1261/rna.056838.116 |
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author | Borchardt, Erin K. Meganck, Rita M. Vincent, Heather A. Ball, Christopher B. Ramos, Silvia B.V. Moorman, Nathaniel J. Marzluff, William F. Asokan, Aravind |
author_facet | Borchardt, Erin K. Meganck, Rita M. Vincent, Heather A. Ball, Christopher B. Ramos, Silvia B.V. Moorman, Nathaniel J. Marzluff, William F. Asokan, Aravind |
author_sort | Borchardt, Erin K. |
collection | PubMed |
description | Circular RNAs (circRNAs) are highly stable, covalently closed RNAs that are regulated in a spatiotemporal manner and whose functions are largely unknown. These molecules have the potential to be incorporated into engineered systems with broad technological implications. Here we describe a switch for inducing back-splicing of an engineered circRNA that relies on the CRISPR endoribonuclease, Csy4, as an activator of circularization. The endoribonuclease activity and 3′ end-stabilizing properties of Csy4 are particularly suited for this task. Coexpression of Csy4 and the circRNA switch allows for the removal of downstream competitive splice sites and stabilization of the 5′ cleavage product. This subsequently results in back-splicing of the 5′ cleavage product into a circRNA that can translate a reporter protein from an internal ribosomal entry site (IRES). Our platform outlines a straightforward approach toward regulating splicing and could find potential applications in synthetic biology as well as in studying the properties of different circRNAs. |
format | Online Article Text |
id | pubmed-5393173 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-53931732018-05-01 Inducing circular RNA formation using the CRISPR endoribonuclease Csy4 Borchardt, Erin K. Meganck, Rita M. Vincent, Heather A. Ball, Christopher B. Ramos, Silvia B.V. Moorman, Nathaniel J. Marzluff, William F. Asokan, Aravind RNA Report Circular RNAs (circRNAs) are highly stable, covalently closed RNAs that are regulated in a spatiotemporal manner and whose functions are largely unknown. These molecules have the potential to be incorporated into engineered systems with broad technological implications. Here we describe a switch for inducing back-splicing of an engineered circRNA that relies on the CRISPR endoribonuclease, Csy4, as an activator of circularization. The endoribonuclease activity and 3′ end-stabilizing properties of Csy4 are particularly suited for this task. Coexpression of Csy4 and the circRNA switch allows for the removal of downstream competitive splice sites and stabilization of the 5′ cleavage product. This subsequently results in back-splicing of the 5′ cleavage product into a circRNA that can translate a reporter protein from an internal ribosomal entry site (IRES). Our platform outlines a straightforward approach toward regulating splicing and could find potential applications in synthetic biology as well as in studying the properties of different circRNAs. Cold Spring Harbor Laboratory Press 2017-05 /pmc/articles/PMC5393173/ /pubmed/28223408 http://dx.doi.org/10.1261/rna.056838.116 Text en © 2017 Borchardt et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. |
spellingShingle | Report Borchardt, Erin K. Meganck, Rita M. Vincent, Heather A. Ball, Christopher B. Ramos, Silvia B.V. Moorman, Nathaniel J. Marzluff, William F. Asokan, Aravind Inducing circular RNA formation using the CRISPR endoribonuclease Csy4 |
title | Inducing circular RNA formation using the CRISPR endoribonuclease Csy4 |
title_full | Inducing circular RNA formation using the CRISPR endoribonuclease Csy4 |
title_fullStr | Inducing circular RNA formation using the CRISPR endoribonuclease Csy4 |
title_full_unstemmed | Inducing circular RNA formation using the CRISPR endoribonuclease Csy4 |
title_short | Inducing circular RNA formation using the CRISPR endoribonuclease Csy4 |
title_sort | inducing circular rna formation using the crispr endoribonuclease csy4 |
topic | Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5393173/ https://www.ncbi.nlm.nih.gov/pubmed/28223408 http://dx.doi.org/10.1261/rna.056838.116 |
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