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...
Autores principales: | , , , , , |
---|---|
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 |
_version_ | 1783528013972897792 |
---|---|
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. |
format | Online Article Text |
id | pubmed-7192461 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT jacobsenthomas tunableselfcleavingribozymesformodulatinggeneexpressionineukaryoticsystems AT yigloria tunableselfcleavingribozymesformodulatinggeneexpressionineukaryoticsystems AT alasafenhadel tunableselfcleavingribozymesformodulatinggeneexpressionineukaryoticsystems AT jermusykashleya tunableselfcleavingribozymesformodulatinggeneexpressionineukaryoticsystems AT beiselchasel tunableselfcleavingribozymesformodulatinggeneexpressionineukaryoticsystems AT reevesgregoryt tunableselfcleavingribozymesformodulatinggeneexpressionineukaryoticsystems |