Intellectual Properties and Commercialisation

The SBRC Nottingham has developed new synthetic biology tools, microbial chassis and resources which are being utilised internationally by industry as well as in academia.

Our intention is to maximise the research, environmental social and economic impacts from the Research Councils' funding that underpins the Centre.

Ambition

Wherever practicable, research tools we develop are made widely available to industry and academia through appropriate non-exclusive research and / or commercialisation licences.

The distribution of many of the SBRC's plasmid vectors is managed through Chain Biotech Ltd (https://chainbiotools.com/product/pmtl80000-vector-series-2/)

To discuss your licensing needs, please contact Dr Alan Burbidge, SBRC Nottingham Centre Manager: alan.burbidge@nottingham.ac.uk

 

Patented Technologies

Synthetic Operon Construction in Clostridia Allele Coupled Exchange (ACE)

WO 2009/101400

An homologous recombination system that avoids the need for a heterologous reporter system and which enables large tracts of DNA to be inserted into a host bacterium such as Clostridium acetylbutylicum.

Academic Publication

  • Integration of DNA into bacterial chromosomes from plasmids without a counter-selection marker. John T. Heap, Muhammad Ehsaan, Clare M. Cooksley, Yen-Kuan Ng, Stephen T. Cartman, Klaus Winzer, and Nigel P. Minton* Nucleic Acids Res. (2012); 40(8): e59.
  • Expanding the repertoire of gene tools for precise manipulation of the Clostridium difficile genome: allelic exchange using pyrE alleles. Ng YK, Ehsaan M, Philip S, Collery MM, Janoir C, Collignon A, Cartman ST and Minton NP, (2013). PloS one. 8(2), e56051.

A Negative/Counter Selection Marker for Use in Clostridia: Cod A Selectable Marker

WO 2010/084349

The cytosine deaminase gene (codA) of Escherichia coli as a heterologous counter-selection marker for genetic manipulation of wild-type C. difficile strains. CodA not only converts cytosine to uracil but also converts the innocuous pyrimidine analog 5-fluorocytosine (FC) into the highly toxic 5-fluorouracil (FU) leading to incorporation of fluorinated nucleotides into DNA and RNA. It is this latter activity which allows CodA to be an effective counter-selection marker.

Academic Publication

  • Precise manipulation of the Clostridium difficile chromosome reveals a lack of association between tcdC genotype and toxin production. Cartman ST, Kelly ML, Heeg D, Heap JT, Minton NP (2012). Applied Environmental Microbiology 78: 4683–90

Transposon Delivery System for Clostridia (vector) TraDis Transposon System

WO 2013/144653

A plasmid-based transposon system in which Clostridial cells transformed with the plasmid results in cut and paste integration of the transposon into chromosomal DNA with a selectable marker. Loss of the plasmid in non-permissive conditions (using inducible promoters) prevents further transposition and facilitates selection of transposon-carrying colonies and the creation of transposon libraries. The patent facilitates TraDIS (Langridge et al. (2009) Genome Res. 19: 2308-2316.

Orthogonal Bacterial Expression System

WO 2013/144647

An expression system exploiting Group 5 RNA polymerase sigma factors for example the TcdR Sigma factor from Clostridium difficile. Such factors are poorly recognised in bacteria lacking Group 5 RNA Polymerase. An example of an orthogonal construct would be TcdR operably linked to a promoter recognised by TcdR and a heterologous DNA sequence in a bacterial host lacking any Group 5 RNA Polymerase and as such give constitutive expression of any TcdR-promoter linked DNA sequence.

ClosTron

WO2009101400 - METHODS

A type II intron system based on TargeTron which enables gene knock-outs through intron insertion in Clostridia. Visit ClosTron at http://clostron.com/

Academic Publication

  • The ClosTron: A universal gene knock-out system for the genus Clostridium. Heap, J.T., Pennington, O.J., Cartman, S.T., Carter, G.P. and Minton, N.P., 2007. Journal of Microbiological Methods. 70(3), 452-464.
  • The ClosTron: Mutagenesis in Clostridium refined and streamlined. Heap JT, Kuehne SA, Ehsaan M, Cartman ST, Cooksley CM, Scott JC, Minton NP. J Microbiol Methods. 2010 Jan;80(1):49-55.

RiboCas

WO2020157483 - GENETIC CONSTRUCT

A universal CRISPR-Cas9 mutagenesis system for broad application in Clostridium species. Many CRISPR-Cas9 systems suffer from toxicity issues in both E. coli and Clostridium cells as a consequence of poor regulatory control of Cas9. As such, many systems utilise nCas9, a mutated derivative which possesses only nickase activity, thus lacking powerful selection capacity. In the RiboCas system, Cas9 has been placed under the control of the constitutive promoter Pfdx which itself, has been linked with a theophylline-responsive riboswitch. The result of which is a tightly repressed Cas9 mutagenesis system, which can be easily activated by inclusion of inducer compound. Doing so, circumvents the issues relating to Cas9 toxicity, thus permitting the use of wild-type Cas9 for mutagenesis studies in Clostridium spp.

RiboLac

WO2020157487 - GENETIC CONSTRUCT AND USES THEREOF

RiboLac system for controlled conditional gene expression in Clostridia

The RiboLac technology describes a two-tiered system for the controlled and conditional expression of target genes in Clostridia. The main advantage of the system is that it acts at both transcriptional and translational levels, allowing high expression levels in the presence of two inducers and very low leakage in their absence. 

At the transcriptional level, the activator BgaR requires the presence of lactose to “switch on” the transcription of the target gene. At the translational level, a riboswitch is placed upstream of the gene encoding BgaR and prevents its translation in the absence of theophylline. As a result, target gene activity is dependent on the presence and concentration of both inducers, lactose and theophylline, simultaneously: theophylline allows the translation of the BgaR activator which, in the presence of lactose, induces the expression of the target gene. 

The system thus allows for tight regulation of gene expression under fermentation conditions and, although both inducers do occur naturally, environmental induction would be deemed negligible. The SBRC has used this system to demonstrate very tight control of sporulation in Clostridia by targeting the master regulator for sporulation, Spo0A.

Academic Publication

  • RiboCas: A Universal CRISPR-Based Editing Tool for Clostridium. Cañadas IC, Groothuis D, Maria Z, Raquel R and Minton NPM. ACS Synthetic Biology. 2019 8 (6), 1379-1390 DOI: 10.1021/acssynbio.9b00075