Dual transcriptional-translational cascade permits cellular level tuneable expression control.

Rosa Morra, Jayendra Shankar, Christopher Robinson, Samantha Halliwell, Lisa Butler, Mathew Upton, Sam Hay, Jason Micklefield, Neil Dixon

    Research output: Contribution to journalArticlepeer-review

    Abstract

    The ability to induce gene expression in a small molecule dependent manner has led to many applications in target discovery, functional elucidation and bio-production. To date these applications have relied on a limited set of protein-based control mechanisms operating at the level of transcription initiation. The discovery, design and reengineering of riboswitches offer an alternative means by which to control gene expression. Here we report the development and characterization of a novel tunable recombinant expression system, termed RiboTite, which operates at both the transcriptional and translational level. Using standard inducible promoters and orthogonal riboswitches, a multi-layered modular genetic control circuit was developed to control the expression of both bacteriophage T7 RNA polymerase and recombinant gene(s) of interest. The system was benchmarked against a number of commonly used E. coli expression systems, and shows tight basal control, precise analogue tunability of gene expression at the cellular level, dose-dependent regulation of protein production rates over extended growth periods and enhanced cell viability. This novel system expands the number of E. coli expression systems for use in recombinant protein production and represents a major performance enhancement over and above the most widely used expression systems.
    Original languageEnglish
    Article numbere21
    JournalNucleic acids research
    Volume44
    Issue number3
    Early online date23 Sept 2015
    DOIs
    Publication statusPublished - 18 Feb 2016

    Research Beacons, Institutes and Platforms

    • Manchester Institute of Biotechnology

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