14TSB_SynBio Automated Gene Assembly From Codons to Complete Genes and Pathways

The ability to assemble genes from smaller DNA fragments or generate combinatorial libraries of genetic mutations is fundamental to the field of synthetic biology. Different methodologies exist for their fabrication but enzymatic assembly produces consistently high quality products compared to other methods which is essential for downstream processes for improved characteristics (whether this is for improved strain phenotype or an improved protein function/yield). The potential to achieve this in a high throughput, automatable fashion would be a powerful and valuable tool for commercialisation in the synthetic biology arena. In this respect, we have previously developed a technology that can create random stretches of gene fragments using enzymatic additions of codon triplets. We now wish to investigate further procedures to assemble diversified gene fragments, in particular antibody variable loops, into whole genes in an automated fashion. Imperial College is automating assembly methods that would be enabling for the project by bridging the scale from codons to genes and pathways, thereby potentially the opening up new market areas.

The ability to assemble genes from smaller DNA fragments or generate combinatorial libraries of genetic mutations is fundamental to the field of synthetic biology. Different methodologies exist for their fabrication but enzymatic assembly produces consistently high quality products compared to other methods which is essential for downstream processes for improved characteristics (whether this is for improved strain phenotype or an improved protein function/yield). The potential to achieve this in a high throughput, automatable fashion would be a powerful and valuable tool for commercialisation in the synthetic biology arena. In this respect, we have previously developed a technology that can create random stretches of gene fragments using enzymatic additions of codon triplets. We now wish to investigate further procedures to assemble diversified gene fragments, in particular antibody variable loops, into whole genes in an automated fashion. Imperial College is automating assembly methods that would be enabling for the project by bridging the scale from codons to genes and pathways, thereby potentially the opening up new market areas.

The beneficiaries of this project can be defined as the researchers involved, and both the synthetic biology and academic communities as well as industry.
- Measurable impacts from this will include research output, interactions and presentations by the PI and PDRA at meetings both internally, externally and internationally, as well as the future placement of the PDRA involved.
The broader academic synthetic biology community will stand to benefit from this research.
- Measurable impacts from this research will be the development of new platform approaches that lead to the integration of codon diversification with DNA assembly of larger fragments.

Enzyme engineering is currently an enormous industry that underpins many biotech applications. The emergence of synthetic biology and its ability to more rapidly develop biological systems and new pathways for the production of fine chemicals in a sustainable manner will only increase the demand for new protein functionality and specificity. The technology that we present in this grant has the potential to be a disruptive technology in this space and radically alter the way that people perceive for evolving new functionality and specificity. Its application is not specific to a single domain but can be envisioned equally in antibody engineering for sensors, diagnostics and thereapeutics as well as enzymatic pathways for production of value chemicals.
- Measurable impacts will be implementation of the new platform within Isogenica and an ongoing collaboration between Imperial College and Isogenica.

The ethos of the Synthetic Biology community and CSynBI is to include public dialogue and outreach at every stage of research from tools to applications.
- Measurable impacts will be any future engagement activities undertaken during the lifetime of the grant.

Dissemination of results is a key feature of scientific research and the scientific endeavour.
- Measurable impacts will be publications and presentations during the lifetime of the grant and the following 12 months