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A positive readout single transcript reporter for site-specific mRNA cleavage
Cleavage of mRNA molecules causes their rapid degradation, thereby playing an important role in regulation of gene expression and host genome defense from viruses and transposons in bacterial and eukaryotic cells. Current negative-readout, and repressor-based positive-readout reporters of mRNA degra...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
PeerJ Inc.
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5522606/ https://www.ncbi.nlm.nih.gov/pubmed/28740759 http://dx.doi.org/10.7717/peerj.3602 |
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author | Kandul, Nikolay Guo, Ming Hay, Bruce A. |
author_facet | Kandul, Nikolay Guo, Ming Hay, Bruce A. |
author_sort | Kandul, Nikolay |
collection | PubMed |
description | Cleavage of mRNA molecules causes their rapid degradation, thereby playing an important role in regulation of gene expression and host genome defense from viruses and transposons in bacterial and eukaryotic cells. Current negative-readout, and repressor-based positive-readout reporters of mRNA degradation have limitations. Here we report the development of a single transcript that acts as a positive reporter of mRNA cleavage. We show that placement of bacterial CopT and CopA hairpins into the 5′ UTR and 3′ UTR of an mRNA results in inhibition of translation of the intervening coding sequence in Drosophila. An internal poly(A) tract inserted downstream of the coding sequence stabilizes transcripts cut within the 3′ UTR. When these components are combined in a transcript in which targets sites for RNA cleavage are placed between the poly(A) tract and CopA, cleavage results in translational activation, providing a single transcript-based method of sensing mRNA cleavage with a positive readout. |
format | Online Article Text |
id | pubmed-5522606 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | PeerJ Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-55226062017-07-24 A positive readout single transcript reporter for site-specific mRNA cleavage Kandul, Nikolay Guo, Ming Hay, Bruce A. PeerJ Bioengineering Cleavage of mRNA molecules causes their rapid degradation, thereby playing an important role in regulation of gene expression and host genome defense from viruses and transposons in bacterial and eukaryotic cells. Current negative-readout, and repressor-based positive-readout reporters of mRNA degradation have limitations. Here we report the development of a single transcript that acts as a positive reporter of mRNA cleavage. We show that placement of bacterial CopT and CopA hairpins into the 5′ UTR and 3′ UTR of an mRNA results in inhibition of translation of the intervening coding sequence in Drosophila. An internal poly(A) tract inserted downstream of the coding sequence stabilizes transcripts cut within the 3′ UTR. When these components are combined in a transcript in which targets sites for RNA cleavage are placed between the poly(A) tract and CopA, cleavage results in translational activation, providing a single transcript-based method of sensing mRNA cleavage with a positive readout. PeerJ Inc. 2017-07-20 /pmc/articles/PMC5522606/ /pubmed/28740759 http://dx.doi.org/10.7717/peerj.3602 Text en ©2017 Kandul et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited. |
spellingShingle | Bioengineering Kandul, Nikolay Guo, Ming Hay, Bruce A. A positive readout single transcript reporter for site-specific mRNA cleavage |
title | A positive readout single transcript reporter for site-specific mRNA cleavage |
title_full | A positive readout single transcript reporter for site-specific mRNA cleavage |
title_fullStr | A positive readout single transcript reporter for site-specific mRNA cleavage |
title_full_unstemmed | A positive readout single transcript reporter for site-specific mRNA cleavage |
title_short | A positive readout single transcript reporter for site-specific mRNA cleavage |
title_sort | positive readout single transcript reporter for site-specific mrna cleavage |
topic | Bioengineering |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5522606/ https://www.ncbi.nlm.nih.gov/pubmed/28740759 http://dx.doi.org/10.7717/peerj.3602 |
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