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Dynamic allocation of orthogonal ribosomes facilitates uncoupling of co-expressed genes

Alexander P.S. Darlington, Juhyun Kim, José I. Jiménez, Declan G. Bates
doi: https://doi.org/10.1101/138362
Alexander P.S. Darlington
1Warwick Integrative Synthetic Biology Centre, School of Engineering, University of Warwick, Coventry, CV4 7AL, UK
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Juhyun Kim
2Faculty of Health and Medical Sciences, University of Surrey, Guildford, GU2 7XH, UK
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José I. Jiménez
2Faculty of Health and Medical Sciences, University of Surrey, Guildford, GU2 7XH, UK
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  • For correspondence: j.jimenez@surrey.ac.uk d.bates@warwick.ac.uk
Declan G. Bates
1Warwick Integrative Synthetic Biology Centre, School of Engineering, University of Warwick, Coventry, CV4 7AL, UK
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  • For correspondence: j.jimenez@surrey.ac.uk d.bates@warwick.ac.uk
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Abstract

Introduction of synthetic circuits into host microbes creates competition between circuit and host genes for shared cellular resources, such as RNA polymerases and ribosomes. This can lead to the emergence of unwanted coupling between the expression of different genes, complicating circuit design and potentially leading to circuit failure. Here we demonstrate the ability of orthogonal ribosomes to alleviate the effects of this resource competition. We partition the ribosome pool by expressing an engineered 16S RNA with altered specificity, and use this division of specificity to build simple resource allocators which reduce the level of ribosome-mediated gene coupling. We then design and implement a dynamic resource allocation controller, which acts to increase orthogonal ribosome production as the demand for translational resources by a synthetic circuit increases. Our results highlight the potential of dynamic translational resource allocation as a means of minimising the impact of cellular limitations on the function of synthetic circuitry.

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Posted May 16, 2017.
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Dynamic allocation of orthogonal ribosomes facilitates uncoupling of co-expressed genes
Alexander P.S. Darlington, Juhyun Kim, José I. Jiménez, Declan G. Bates
bioRxiv 138362; doi: https://doi.org/10.1101/138362
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Dynamic allocation of orthogonal ribosomes facilitates uncoupling of co-expressed genes
Alexander P.S. Darlington, Juhyun Kim, José I. Jiménez, Declan G. Bates
bioRxiv 138362; doi: https://doi.org/10.1101/138362

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