The genetic insulator RiboJ increases expression of insulated genes
| Autor: | Andrew D. Halleran, John P. Marken, Callan E. Monette, Ethan M. Jones, Sudip Paudel, Margaret S. Saha, Kalen P. Clifton, Lidia Epp |
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| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Environmental Engineering Characterization 0206 medical engineering Biomedical Engineering Insulator (electricity) 02 engineering and technology Computational biology Biology 01 natural sciences 03 medical and health sciences Synthetic biology Ribozyme 010608 biotechnology Quantitative assessment Letters to the Editor Molecular Biology Gene lcsh:QH301-705.5 030304 developmental biology RiboJ 0303 health sciences Reporter gene Digital droplet PCR Promoter Cell Biology Genetic circuit 030104 developmental biology lcsh:Biology (General) Insulation biology.protein Protein abundance Developmental biology 020602 bioinformatics |
| Zdroj: | Journal of Biological Engineering Journal of Biological Engineering, Vol 12, Iss 1, Pp 1-6 (2018) |
| ISSN: | 1754-1611 |
| Popis: | A primary objective of synthetic biology is the construction of genetic circuits with behaviors that can be predicted based on the properties of the constituent genetic parts from which they are built. However a significant issue in the construction of synthetic genetic circuits is a phenomenon known as context dependence in which the behavior of a given part changes depending on the choice of adjacent or nearby parts. Interactions between parts compromise the modularity of the circuit, impeding the implementation of predictable genetic constructs. To address this issue, investigators have devised genetic insulators that prevent these unintended context-dependent interactions between neighboring parts. One of the most commonly used insulators in bacterial systems is the self-cleaving ribozyme RiboJ. Despite its utility as an insulator, there has been no systematic quantitative assessment of the effect of RiboJ on the expression level of downstream genetic parts. Here, we characterized the impact of insulation with RiboJ on expression of a reporter gene driven by a promoter from a library of 24 frequently employed constitutive promoters in an Escherichia coli model system. We show that, depending on the strength of the promoter, insulation with RiboJ increased protein abundance between twofold and tenfold and increased transcript abundance by an average of twofold. This result demonstrates that genetic insulators in E. coli can impact the expression of downstream genes, information that is essential for the design of predictable genetic circuits and constructs. Electronic supplementary material The online version of this article (10.1186/s13036-018-0115-6) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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