Tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems

Advancements in the field of synthetic biology have been possible due to the development of genetic tools that are able to regulate gene expression. However, the current toolbox of gene regulatory tools for eukaryotic systems have been outpaced by those developed for simple, single-celled systems. H...

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Autores principales: Jacobsen, Thomas, Yi, Gloria, Al Asafen, Hadel, Jermusyk, Ashley A., Beisel, Chase L., Reeves, Gregory T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7192461/
https://www.ncbi.nlm.nih.gov/pubmed/32352996
http://dx.doi.org/10.1371/journal.pone.0232046
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author Jacobsen, Thomas
Yi, Gloria
Al Asafen, Hadel
Jermusyk, Ashley A.
Beisel, Chase L.
Reeves, Gregory T.
author_facet Jacobsen, Thomas
Yi, Gloria
Al Asafen, Hadel
Jermusyk, Ashley A.
Beisel, Chase L.
Reeves, Gregory T.
author_sort Jacobsen, Thomas
collection PubMed
description Advancements in the field of synthetic biology have been possible due to the development of genetic tools that are able to regulate gene expression. However, the current toolbox of gene regulatory tools for eukaryotic systems have been outpaced by those developed for simple, single-celled systems. Here, we engineered a set of gene regulatory tools by combining self-cleaving ribozymes with various upstream competing sequences that were designed to disrupt ribozyme self-cleavage. As a proof-of-concept, we were able to modulate GFP expression in mammalian cells, and then showed the feasibility of these tools in Drosophila embryos. For each system, the fold-reduction of gene expression was influenced by the location of the self-cleaving ribozyme/upstream competing sequence (i.e. 5′ vs. 3′ untranslated region) and the competing sequence used. Together, this work provides a set of genetic tools that can be used to tune gene expression across various eukaryotic systems.
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spelling pubmed-71924612020-05-11 Tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems Jacobsen, Thomas Yi, Gloria Al Asafen, Hadel Jermusyk, Ashley A. Beisel, Chase L. Reeves, Gregory T. PLoS One Research Article Advancements in the field of synthetic biology have been possible due to the development of genetic tools that are able to regulate gene expression. However, the current toolbox of gene regulatory tools for eukaryotic systems have been outpaced by those developed for simple, single-celled systems. Here, we engineered a set of gene regulatory tools by combining self-cleaving ribozymes with various upstream competing sequences that were designed to disrupt ribozyme self-cleavage. As a proof-of-concept, we were able to modulate GFP expression in mammalian cells, and then showed the feasibility of these tools in Drosophila embryos. For each system, the fold-reduction of gene expression was influenced by the location of the self-cleaving ribozyme/upstream competing sequence (i.e. 5′ vs. 3′ untranslated region) and the competing sequence used. Together, this work provides a set of genetic tools that can be used to tune gene expression across various eukaryotic systems. Public Library of Science 2020-04-30 /pmc/articles/PMC7192461/ /pubmed/32352996 http://dx.doi.org/10.1371/journal.pone.0232046 Text en © 2020 Jacobsen 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, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Jacobsen, Thomas
Yi, Gloria
Al Asafen, Hadel
Jermusyk, Ashley A.
Beisel, Chase L.
Reeves, Gregory T.
Tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems
title Tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems
title_full Tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems
title_fullStr Tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems
title_full_unstemmed Tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems
title_short Tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems
title_sort tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7192461/
https://www.ncbi.nlm.nih.gov/pubmed/32352996
http://dx.doi.org/10.1371/journal.pone.0232046
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