The BUILD platform is responsible for the production of catalytic, regulatory and structural parts, assembly of these parts into pathways, circuits and scaffolds, and engineering of the chassis for efficient expression and high product yields.
PARTS production – The challenge is to create diverse and novel parts libraries, with rapid characterisation, structural elucidation and engineering of bio-catalytic parts. Assembly of directed evolution libraries via DNA synthesis (SpeedyGenes); Automated part preparation (PCR/digestion/clean-up); High throughput QC of parts via capillary electrophoresis; collection of parts depository. Novel proteins derived from the DESIGN platform will be built and screened for biocatalytic activity for targeted pathways. Directed evolution will be employed to engineer novel, improved or altered catalytic activity for biocatalysts.
Biosensors: Bespoke orthogonal riboswitches and other biosensors are being engineered to aid rapid identification of biocatalytic parts activity, and signalling and light-responsive transcriptional regulators will provide temporal control.
DEVICES– Pathways tested in silico in the DESIGN platform are assembled (e.g. Gibson, In-Fusion, Ligase cycling reaction) from synthesised or PCR-based DNA building blocks encoding regulatory and biocatalytic parts to generate combinatorial libraries for screening (SpeedyGenes). HTP assembly methods for large (>10kb) and complex multi-modular pathways are being employed to identify optimal refactored pathways and end products analysed by high throughput screening and state-of-the-art MS techniques in the TEST platform.
SYSTEMS– HTP engineering of optimised chassis includes the assembly and development of industrially relevant, characterised production strain library (bacteria/yeast), the creation of rationally DESIGNed strain modifications by recombinant genetics and cutting-edge genome editing technologies (CRISPR/Cas9), pathway compartmentalisation for reduced intermediate toxicity and increase pathway flux and modifications to the cell envelope for increased stability and product secretion.
Dr Andrew Currin – Directed Evolution of Bio-Parts (Andrew.Currin@manchester.ac.uk)
Dr Mark Dunstan – High Throughput Robotic Platforms (Mark.Dunstan@manchester.ac.uk)
Dr Adrian Jervis – Pathway Engineering (Adrian.Jervis@manchester.ac.uk)
Dr Christopher Robinson – Regulatory Toolkit and Chassis Engineering (Christopher.Robinson@manchester.ac.uk)