Suess, Beatrix ; Hanson, Shane ; Berens, Christian ; Fink, Barbara ; Schroeder, Renée ; Hillen, Wolfgang (2003)
Conditional gene expression by controlling translation with tetracycline-binding aptamers.
In: Nucleic acids research, 31 (7)
Artikel, Bibliographie
Kurzbeschreibung (Abstract)
We present a conditional gene expression system in Saccharomyces cerevisiae which exploits direct RNA-metabolite interactions as a mechanism of genetic control. We inserted preselected tetracycline (tc) binding aptamers into the 5'-UTR of a GFP encoding mRNA. While aptamer insertion generally reduces GFP expression, one group of aptamers displayed an additional, up to 6-fold, decrease in fluorescence upon tc addition. Regulation is observed for aptamers inserted cap-proximal or near the start codon, but is more pronounced from the latter position. Increasing the thermodynamic stability of the aptamer augments regulation but reduces expression of GFP. Decreasing the stability leads to the opposite effect. We defined nucleotides which influence the regulatory properties of the aptamer. Exchanging a nucleotide probably involved in tc binding only influences regulation, while mutations at another position alter expression in the absence of tc, without affecting regulation. Thus, we have developed and characterized a regulatory system which is easy to establish and controlled by a non-toxic, small ligand with good cell permeability.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2003 |
| Autor(en): | Suess, Beatrix ; Hanson, Shane ; Berens, Christian ; Fink, Barbara ; Schroeder, Renée ; Hillen, Wolfgang |
| Art des Eintrags: | Bibliographie |
| Titel: | Conditional gene expression by controlling translation with tetracycline-binding aptamers. |
| Sprache: | Englisch |
| Publikationsjahr: | 2003 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Nucleic acids research |
| Jahrgang/Volume einer Zeitschrift: | 31 |
| (Heft-)Nummer: | 7 |
| Kurzbeschreibung (Abstract): | We present a conditional gene expression system in Saccharomyces cerevisiae which exploits direct RNA-metabolite interactions as a mechanism of genetic control. We inserted preselected tetracycline (tc) binding aptamers into the 5'-UTR of a GFP encoding mRNA. While aptamer insertion generally reduces GFP expression, one group of aptamers displayed an additional, up to 6-fold, decrease in fluorescence upon tc addition. Regulation is observed for aptamers inserted cap-proximal or near the start codon, but is more pronounced from the latter position. Increasing the thermodynamic stability of the aptamer augments regulation but reduces expression of GFP. Decreasing the stability leads to the opposite effect. We defined nucleotides which influence the regulatory properties of the aptamer. Exchanging a nucleotide probably involved in tc binding only influences regulation, while mutations at another position alter expression in the absence of tc, without affecting regulation. Thus, we have developed and characterized a regulatory system which is easy to establish and controlled by a non-toxic, small ligand with good cell permeability. |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie > Synthetic Genetic Circuits (2020 umbenannt in "Synthetic RNA biology") 10 Fachbereich Biologie |
| Hinterlegungsdatum: | 22 Feb 2012 10:41 |
| Letzte Änderung: | 05 Mär 2013 09:59 |
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