Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Borchardt, Erin K., Meganck, Rita M., Vincent, Heather A., Ball, Christopher B., Ramos, Silvia B.V., Moorman, Nathaniel J., Marzluff, William F., Asokan, Aravind
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2017
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
_version_ 1783229553207934976
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
work_keys_str_mv AT borchardterink inducingcircularrnaformationusingthecrisprendoribonucleasecsy4
AT meganckritam inducingcircularrnaformationusingthecrisprendoribonucleasecsy4
AT vincentheathera inducingcircularrnaformationusingthecrisprendoribonucleasecsy4
AT ballchristopherb inducingcircularrnaformationusingthecrisprendoribonucleasecsy4
AT ramossilviabv inducingcircularrnaformationusingthecrisprendoribonucleasecsy4
AT moormannathanielj inducingcircularrnaformationusingthecrisprendoribonucleasecsy4
AT marzluffwilliamf inducingcircularrnaformationusingthecrisprendoribonucleasecsy4
AT asokanaravind inducingcircularrnaformationusingthecrisprendoribonucleasecsy4