Publikation: Neomycin-dependent hammerhead ribozymes for the direct control of gene expression in Saccharomyces cerevisiae
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2019
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Methods. 2019, 161, pp. 35-40. ISSN 1046-2023. eISSN 1095-9130. Available under: doi: 10.1016/j.ymeth.2019.01.004
Zusammenfassung
Hammerhead ribozyme-based RNA switches have been proven to be powerful tools for conditional gene regulation in various organisms. We present neomycin-dependent hammerhead ribozymes (HHR) that influence gene expression in a ligand- and dose-dependent manner in S. cerevisiae. We utilized a novel design of fusing the aptamer domain to the HHR enabling for the first time the identification of genetic ON- and OFF-switches within the same library. For this purpose a neomycin aptamer was fused to stem 1 of a type 3 hammerhead ribozyme via an addressable three-way junction that shows high flexibility at the connection site. An in vivo screening approach identified sequences that allow to induce or repress gene expression 2- to 3-fold in response to neomycin addition. The ribozyme switches operate at neomycin concentrations that show no toxic effect on cell growth and pose powerful genetic tools to study and modulate cellular function in yeast.
Zusammenfassung in einer weiteren Sprache
Fachgebiet (DDC)
540 Chemie
Schlagwörter
Hammerhead ribozyme; Aptazyme; Neomycin; Gene expression regulation
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FINKE, Monika, Julia STIFEL, Stefan G. KREFT, Elke DEUERLING, Jörg S. HARTIG, 2019. Neomycin-dependent hammerhead ribozymes for the direct control of gene expression in Saccharomyces cerevisiae. In: Methods. 2019, 161, pp. 35-40. ISSN 1046-2023. eISSN 1095-9130. Available under: doi: 10.1016/j.ymeth.2019.01.004BibTex
@article{Finke2019-05Neomy-44869,
year={2019},
doi={10.1016/j.ymeth.2019.01.004},
title={Neomycin-dependent hammerhead ribozymes for the direct control of gene expression in Saccharomyces cerevisiae},
volume={161},
issn={1046-2023},
journal={Methods},
pages={35--40},
author={Finke, Monika and Stifel, Julia and Kreft, Stefan G. and Deuerling, Elke and Hartig, Jörg S.}
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