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

by Beisel, Chase L. , Al Asafen, Hadel , Jacobsen, Thomas , Reeves, Gregory T. , Jermusyk, Ashley A. , Yi, Gloria

in 3' Untranslated regions / Animals / Bacteria / Binding sites / Biology and Life Sciences / Drosophila - genetics / Embryos / Engineering / Engineers / Eukaryota - genetics / Eukaryota - metabolism / Eukaryotic Cells - metabolism / Fruit flies / Gene expression / Gene Expression - genetics / Gene Expression - physiology / Gene Expression Regulation - genetics / Gene Expression Regulation - physiology / Genetic Engineering - methods / Insects / Mammalian cells / Nucleic Acid Conformation / Physical Sciences / Plasmids / Proof of Concept Study / Protein synthesis / Proteins / Research and Analysis Methods / Ribozymes / RNA, Catalytic - genetics / RNA, Catalytic - physiology / RNA, Messenger - metabolism / Synthetic Biology - methods / Transcription factors

2020

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

by Beisel, Chase L. , Al Asafen, Hadel , Jacobsen, Thomas , Reeves, Gregory T. , Jermusyk, Ashley A. , Yi, Gloria

2020

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

by Beisel, Chase L. , Al Asafen, Hadel , Jacobsen, Thomas , Reeves, Gregory T. , Jermusyk, Ashley A. , Yi, Gloria

2020

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Journal Article

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

Beisel, Chase L.,

Al Asafen, Hadel,

Jacobsen, Thomas,

Reeves, Gregory T.,

Jermusyk, Ashley A.,

Yi, Gloria

2020

Overview

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' vs. 3' untranslated region) 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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Publisher

Public Library of Science,Public Library of Science (PLoS)

Subject

3' Untranslated regions

/ Animals

/ Bacteria

/ Binding sites

/ Biology and Life Sciences

/ Drosophila - genetics

/ Embryos