Enzymatic gene synthesis

Lead Participant: Evonetix Ltd

Abstract

"The revolution generated by next-generation sequencing platforms led to a massive increase in DNA sequence information becoming available over the past ten years. There now exists an unprecedented opportunity to engineer metabolic pathways and organisms, improve industrial processes, create new processes, engineer genomes with new and improved traits and use DNA as a medium for digital data storage. All the foregoing will have a fundamental impact on science and industry, and potentially on the quality of life for millions of individuals. This impending area of research and commercial endeavour has been termed ""synthetic biology"".

However, the current inability to produce vast amounts of accurate long, DNA molecules at low cost is limiting the growth of the synthetic biology industry. Current approaches to DNA synthesis involve the generation of oligonucleotides, mostly using phosphoramidite chemistry, followed by annealing and assembly in pools, to generate double-stranded DNA. This is most successful at DNA lengths up to 3 kilobases and less, where accumulated errors do not require the time-consuming and expensive correction of sequences that becomes essential at greater lengths. In addition, current platforms, based on ink-jet and other physical separation modalities, are extremely limited in their ability to secure scaled DNA manufacture. More than 1 billion bases of double-stranded DNA have been synthesised at hugely variable pricing depending on the requirement for post-synthesis correction. Moreover, delivery times for such syntheses have been highly extended and thus many projects remain unfinished.

Evonetix, a Cambridge-based start-up, aims to revolutionise _de novo_ gene synthesis. We are developing a highly disruptive DNA/gene synthesis platform to address the increasing demand for accurate synthetic DNA at low cost.

We believe the use of enzymatic oligonucleotide synthesis, which operates under milder aqueous conditions compared to phosphoramidite chemistry, will provide a significant commercial advantage for _de novo_ DNA/genes synthesis with our platform; it will achieve this by reducing cost, being more environmentally friendly and further streamlining the industrialisation of high-fidelity DNA synthesis.

Enzymes with terminal deoxynucleotidyl transferase/synthetase activities are known to extend single-stranded DNA without the need for a template strand. This unique ability to produce DNA _de novo_ makes these enzymes highly valuable biological tools for the production of synthetic DNA.

The aim of our project is to develop novel, modified nucleotides and genetically engineered enzymes with terminal deoxynucleotidyl transferase/synthetase activities as an alternative to the well-established phosphoramidite chemistry, to synthesise high-quality DNA/genes with our innovative technology."

Lead Participant

Project Cost

Grant Offer

Evonetix Ltd, CAMBRIDGE £933,040 £ 653,128
 

Participant

Durham University, Durham £337,713 £ 337,713

Publications

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