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Tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems

Thomas Jacobsen, Gloria Yi, Hadel A. Asafen, Ashley A. Jermusyk, Chase L. Beisel, View ORCID ProfileGregory T. Reeves
doi: https://doi.org/10.1101/739326
Thomas Jacobsen
1Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695
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Gloria Yi
1Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695
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Hadel A. Asafen
1Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695
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Ashley A. Jermusyk
1Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695
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Chase L. Beisel
1Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695
2Helmholtz Institute for RNA-based Infection Research (HIRI), Helmholtz-Centre for Infection Research (HZI), 97080 Würzburg, Germany
3Medical Faculty, University of Würzburg, 97080 Würzburg, Germany
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  • For correspondence: chase.beisel@helmholtz-hiri.de gtreeves@ncsu.edu
Gregory T. Reeves
1Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695
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  • ORCID record for Gregory T. Reeves
  • For correspondence: chase.beisel@helmholtz-hiri.de gtreeves@ncsu.edu
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ABSTRACT

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’ untranslated region (UTR) vs. 3’UTR) 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.

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Posted August 18, 2019.
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Tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems
Thomas Jacobsen, Gloria Yi, Hadel A. Asafen, Ashley A. Jermusyk, Chase L. Beisel, Gregory T. Reeves
bioRxiv 739326; doi: https://doi.org/10.1101/739326
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Tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems
Thomas Jacobsen, Gloria Yi, Hadel A. Asafen, Ashley A. Jermusyk, Chase L. Beisel, Gregory T. Reeves
bioRxiv 739326; doi: https://doi.org/10.1101/739326

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