Direct and specific chemical control of eukaryotic translation with a synthetic RNA–protein interaction
Sequence-specific RNA–protein interactions, though commonly used in biological systems to regulate translation, are challenging to selectively modulate. Here, we demonstrate the use of a chemically-inducible RNA–protein interaction to regulate eukaryotic translation. By genetically encoding Tet Repr...
| Main Authors: | , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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Oxford University Press (OUP)
2012
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| Online Access: | http://hdl.handle.net/1721.1/73695 https://orcid.org/0000-0002-6250-8796 |
| _version_ | 1811096740852400128 |
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| author | Goldfless, Stephen Jacob Belmont, Brian Joshua de Paz, Alexandra M. Niles, Jacquin |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Goldfless, Stephen Jacob Belmont, Brian Joshua de Paz, Alexandra M. Niles, Jacquin |
| author_sort | Goldfless, Stephen Jacob |
| collection | MIT |
| description | Sequence-specific RNA–protein interactions, though commonly used in biological systems to regulate translation, are challenging to selectively modulate. Here, we demonstrate the use of a chemically-inducible RNA–protein interaction to regulate eukaryotic translation. By genetically encoding Tet Repressor protein (TetR)-binding RNA elements into the 5′-untranslated region (5′-UTR) of an mRNA, translation of a downstream coding sequence is directly controlled by TetR and tetracycline analogs. In endogenous and synthetic 5′-UTR contexts, this system efficiently regulates the expression of multiple target genes, and is sufficiently stringent to distinguish functional from non-functional RNA–TetR interactions. Using a reverse TetR variant, we illustrate the potential for expanding the regulatory properties of the system through protein engineering strategies. |
| first_indexed | 2024-09-23T16:48:18Z |
| format | Article |
| id | mit-1721.1/73695 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:48:18Z |
| publishDate | 2012 |
| publisher | Oxford University Press (OUP) |
| record_format | dspace |
| spelling | mit-1721.1/736952022-09-29T21:35:16Z Direct and specific chemical control of eukaryotic translation with a synthetic RNA–protein interaction Goldfless, Stephen Jacob Belmont, Brian Joshua de Paz, Alexandra M. Niles, Jacquin Massachusetts Institute of Technology. Department of Biological Engineering Goldfless, Stephen Jacob Belmont, Brian Joshua de Paz, Alexandra M. Niles, Jacquin Sequence-specific RNA–protein interactions, though commonly used in biological systems to regulate translation, are challenging to selectively modulate. Here, we demonstrate the use of a chemically-inducible RNA–protein interaction to regulate eukaryotic translation. By genetically encoding Tet Repressor protein (TetR)-binding RNA elements into the 5′-untranslated region (5′-UTR) of an mRNA, translation of a downstream coding sequence is directly controlled by TetR and tetracycline analogs. In endogenous and synthetic 5′-UTR contexts, this system efficiently regulates the expression of multiple target genes, and is sufficiently stringent to distinguish functional from non-functional RNA–TetR interactions. Using a reverse TetR variant, we illustrate the potential for expanding the regulatory properties of the system through protein engineering strategies. National Institutes of Health (U.S.) (Director’s New Innovator Award Program Grant 1DP2OD007124) National Institute of Environmental Health Sciences (Predoctoral Training Grant 5-T32-ES007020) National Science Foundation (U.S.) (Research Experiences for Undergraduates Grant 1005055) 2012-10-10T14:39:58Z 2012-10-10T14:39:58Z 2012-01 2012-01 Article http://purl.org/eprint/type/JournalArticle 0305-1048 1362-4962 http://hdl.handle.net/1721.1/73695 Goldfless, S. J. et al. “Direct and Specific Chemical Control of Eukaryotic Translation with a Synthetic RNA-protein Interaction.” Nucleic Acids Research 40.9 (2012): e64–e64. https://orcid.org/0000-0002-6250-8796 en_US http://dx.doi.org/10.1093/nar/gks028 Nucleic Acids Research Creative Commons Attribution Non-Commercial http://www.creativecommons.org/licenses/by-nc/3.0/ application/pdf Oxford University Press (OUP) Oxford University Press |
| spellingShingle | Goldfless, Stephen Jacob Belmont, Brian Joshua de Paz, Alexandra M. Niles, Jacquin Direct and specific chemical control of eukaryotic translation with a synthetic RNA–protein interaction |
| title | Direct and specific chemical control of eukaryotic translation with a synthetic RNA–protein interaction |
| title_full | Direct and specific chemical control of eukaryotic translation with a synthetic RNA–protein interaction |
| title_fullStr | Direct and specific chemical control of eukaryotic translation with a synthetic RNA–protein interaction |
| title_full_unstemmed | Direct and specific chemical control of eukaryotic translation with a synthetic RNA–protein interaction |
| title_short | Direct and specific chemical control of eukaryotic translation with a synthetic RNA–protein interaction |
| title_sort | direct and specific chemical control of eukaryotic translation with a synthetic rna protein interaction |
| url | http://hdl.handle.net/1721.1/73695 https://orcid.org/0000-0002-6250-8796 |
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