Co-evolution of small molecule responsive riboswitches
Lead Research Organisation:
University of Manchester
Department Name: Chemistry
Abstract
Small molecule modulators of gene expression, which offer rapid, temporal and spatial control of gene expression, would be tremendously valuable tools in biological and biomedical sciences. Indeed there are relatively few examples of small molecule-inducible expression systems that are available for general use. In addition, the ability to control differentially the expression of multiple genes simultaneously, using distinct small molecule inhibitors or inducers, remains extremely limited. In this project we aim to engineer new regulatory systems that function in prokaryotic and eukaryotic cells and respond in a rapid dose-dependent fashion to a wide range of small drug-like molecules. It is envisaged that the ability to control gene expression in response to a wide range of small molecule could lead to many profound applications in pharmaceutical target validation, gene therapy, bioremediation, biosensors, and plant biotechnology.
Technical Summary
Small molecule modulators of gene expression, which offer rapid, temporal and spatial control of gene expression, would be tremendously valuable tools in biological and biomedical sciences. Indeed there are relatively few examples of small molecule-inducible expression systems that are available for general use. In addition, the ability to control differentially the expression of multiple genes simultaneously, using distinct small molecule inhibitors or inducers, remains extremely limited. In this project we aim to engineer new regulatory systems that function in prokaryotic and eukaryotic cells and respond in a rapid dose-dependent fashion to a wide range of small drug-like molecules. It is envisaged that the ability to control gene expression in response to a wide range of small molecule could lead to many profound applications in pharmaceutical target validation, gene therapy, bioremediation, biosensors, and plant biotechnology.
Organisations
Publications
Chen L
(2012)
Probing riboswitch-ligand interactions using thiamine pyrophosphate analogues.
in Organic & biomolecular chemistry
Dixon N
(2010)
Reengineering orthogonally selective riboswitches.
in Proceedings of the National Academy of Sciences of the United States of America
Dixon N
(2012)
Orthogonal riboswitches for tuneable coexpression in bacteria.
in Angewandte Chemie (International ed. in English)
Vincent HA
(2014)
Generation of orthogonally selective bacterial riboswitches by targeted mutagenesis and in vivo screening.
in Methods in molecular biology (Clifton, N.J.)
| Description | We successfully re-engineered the first orthogonal riboswitches, which no longer respond to the natural cellular metabolites, and can be used to activate gene expression in a precise dose-dependent fashion with a variety of synthetic ligands not present within the cell. We have characterised these riboswitches in vivo and in vitro using ITC, SHAPE and X-ray crystallography, producing the first high-resolution structure of an orthogonal riboswitch in complex with a synthetic ligand. A full report detailing all of the outcomes of this research has been submitted to BBSRC and only key publications and patents are listed here. |
| Exploitation Route | This application of the orthogonal riboswitch technology is probably best realised through collaboration with established companies who develop protein expression systems, vectors and expression hosts etc. A patent application has been submitted and development work, supported by BBSRC follow-on-funding, is underway to explore the potential applications of orthogonal riboswitches for protein expression etc. |
| Sectors | Healthcare,Pharmaceuticals and Medical Biotechnology |
| Description | see final report submitted to bbsrc |
| Sector | Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Description | Feasibility and Benchmarking of RiboTite gene expression control technology |
| Amount | £146,000 (GBP) |
| Funding ID | BB/J019089/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 06/2012 |
| End | 09/2013 |
| Title | RIBOSWITCHES |
| Description | The present invention relates to a system comprising a genetic construct a riboswitch operably linked to a regulatory sequence, and a second genetic construct a coding sequence whose expression is capable of being regulated by a gene product of the first construct. Also provided is a genetic construct comprising one or more riboswitches for regulation of gene expression, wherein preferably a spacer sequence is provided downstream of the riboswitch to enhance expression of a coding sequence which is operably linked to a riboswitch. Ligands, kits, methods, host cells and expression systems are also provided. |
| IP Reference | WO2012153142 |
| Protection | Patent application published |
| Year Protection Granted | 2012 |
| Licensed | No |
| Impact | Dixon, N.; Micklefield, J. Riboswitch-regulated transgene expression systems From PCT Int. Appl. (2012), WO 2012153142 A2 20121115. |
| Description | Synthetic Biology Congress, Munich, Germany, October 8-9, 2018. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Other audiences |
| Results and Impact | Presentation about research in the Micklefield group |
| Year(s) Of Engagement Activity | 2014,2018 |