Tools: (Genome editing, Optogenetics, Microfluidics, Protein production, Vectors and gene regulation)
Work is ongoing at Glasgow University to develop a wide range of tools for Synthetic Biology. These range from optogenetic tools to control gene expression using light, DNA-editing enzymes to write digital information into DNA sequences, and using microfluidics and protocell technologies for cell free protein expression and directed evolution strategies.
Optogenetics is an emerging field that combines optical and genetic approaches to non-invasively control cellular events with exquisite spatiotemporal control. The repertoire of light-sensitive proteins used for devising new optogenetic tools is rapidly expanding. Work in Glasgow concentrates on developing LOV domain proteins to detect visible light or UVR8 to detect UV-B. These domains are being used to generate transcriptional regulators, ion channels or other proteins that can be controlled by specific wavelengths of light. One aim is to change the regulation of transport proteins in biological membranes for improved water efficiency in important crop plants.
Enzymes known as site-specific recombinases and transposases carry out cut and paste DNA editing reactions. Site-specific recombination can be used to insert or delete specific DNA sequences in a highly controlled fashion. We are using recombinases to deliver DNA to specific “landing pads” in the genome of biotechnologically important organisms. With our industrial collaborators Ingenza, we are using these methods to manipulate yeast for improved biofuel production. Site-specific recombination can also be used to record digital information in DNA sequences in the form of binary 1s and 0s, and we are using this to develop new biological counters, information storage arrays, and biosensors.
Transposases can be used to insert DNA into the genome of almost any organismThis can be used for gene therapy of other DNA delivery applications. Insertions occur at random locations with possible deleterious effects. We are engineering improved transposase proteins that target insertions to specific sites in the host genome.
Nanoscale devices for Synthetic Biology (Microfluidics and protocells)
Work in this area ranges from development of functional interfaces and integrated microfluidic devices for the investigation of microscale phenomena in biology, using ultrasound to control and integrate synthetic biology processes and parts, production of lipid vesicles to encapsulate cell-free protein expression machinery, and nanophotonics for Synthetic Biology.
Mammalian Protein Expression Systems
One of the major challenges facing the biopharmaceutical industry is to produce sufficient therapeutic proteins in a cost effective way to meet market demand. The level of protein production in mammalian cells is limited by the cell’s capacity to fold and assemble the translated polypeptide chain. This limitation is particularly true for proteins entering the secretory pathway which not only need to fold correctly but also must be post-translationally modified to form the correct disulphide bonds and to be N-link glycosylated. We are currently using a synthetic biology approach to develop efficient engineered mammalian cell factories to maximise the production of novel pharmaceutical proteins.
Synthetic Biology Toolkit for cyanobacteria
Photosynthetic cyanobacteria such as Synechocystis have the potential to be used as biorefineries to produce biofuels, chemical feedstocks or pharmaceuticals powered directly by the energy from sun light. Work is ongoing to produce new plasmid vectors, libraries of regulated promoters, and novel gene delivery methods for improved genetic manipulation of these organisms.