Kötter, Peter ; Weigand, Julia E. ; Meyer, Britta ; Entian, Karl-Dieter ; Suess, Beatrix (2009)
A fast and efficient translational control system for conditional expression of yeast genes.
In: Nucleic acids research, 37 (18)
Artikel, Bibliographie
Kurzbeschreibung (Abstract)
A new artificial regulatory system for essential genes in yeast is described. It prevents translation of target mRNAs upon tetracycline (tc) binding to aptamers introduced into their 5'UTRs. Exploiting direct RNA-ligand interaction renders auxiliary protein factors unnecessary. Therefore, our approach is strain independent and not susceptible to interferences by heterologous expressed regulatory proteins. We use a simple PCR-based strategy, which allows easy tagging of any target gene and the level of gene expression can be adjusted due to various tc aptamer-regulated promoters. As proof of concept, five differently expressed genes were targeted, two of which could not be regulated previously. In all cases, adding tc completely prevented growth and, as shown for Nop14p, rapidly abolished de novo protein synthesis providing a powerful tool for conditional regulation of yeast gene expression.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2009 |
| Autor(en): | Kötter, Peter ; Weigand, Julia E. ; Meyer, Britta ; Entian, Karl-Dieter ; Suess, Beatrix |
| Art des Eintrags: | Bibliographie |
| Titel: | A fast and efficient translational control system for conditional expression of yeast genes. |
| Sprache: | Englisch |
| Publikationsjahr: | 2009 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Nucleic acids research |
| Jahrgang/Volume einer Zeitschrift: | 37 |
| (Heft-)Nummer: | 18 |
| Kurzbeschreibung (Abstract): | A new artificial regulatory system for essential genes in yeast is described. It prevents translation of target mRNAs upon tetracycline (tc) binding to aptamers introduced into their 5'UTRs. Exploiting direct RNA-ligand interaction renders auxiliary protein factors unnecessary. Therefore, our approach is strain independent and not susceptible to interferences by heterologous expressed regulatory proteins. We use a simple PCR-based strategy, which allows easy tagging of any target gene and the level of gene expression can be adjusted due to various tc aptamer-regulated promoters. As proof of concept, five differently expressed genes were targeted, two of which could not be regulated previously. In all cases, adding tc completely prevented growth and, as shown for Nop14p, rapidly abolished de novo protein synthesis providing a powerful tool for conditional regulation of yeast gene expression. |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie 10 Fachbereich Biologie > Synthetic Genetic Circuits (2020 umbenannt in "Synthetic RNA biology") 10 Fachbereich Biologie > RNA Biochemie |
| Hinterlegungsdatum: | 22 Feb 2012 10:27 |
| Letzte Änderung: | 06 Nov 2020 07:21 |
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