Engineering orthogonal split inteins as scalable tools for synthetic biology and biomanufacturing

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An important goal of synthetic biology is the rational design and predictable implementation of synthetic gene circuits using standardised interchangeable parts to program cellular behaviour. However, most of the circuits constructed so far are small-scale systems that have been constructed by costly and inefficient 'trial-and-error' methods with limited parts. A hard truth behind this is that the engineering of complex circuits in living cells is currently limited by the availability of well-characterised and orthogonal genetic regulatory building blocks in addition to the lack of robust wiring methods. Moreover, versatile post-translational regulation tools such as split inteins represent an untapped/underexplored invaluable resource for synthetic biologists. This project aims to develop new enabling tools for scalable gene circuit design via engineering novel orthogonal split intein mediated gene regulation control strategies, including engineering a library of modular and orthogonal genetic logic gates and switches to significantly expand the currently limited toolbox of genetic control components. The outcome will transform the present state of gene network engineering by directly addressing a key bottleneck in the field. I will also develop new efficient automation tools for gene circuit design and diagnosis to achieve rapid, predictable design of large-scale genetic logic circuits with advanced cellular information processing capacity. Such tools have the potential to significantly increase the efficiency and scale, and shorten development time of a typical gene circuit design project from the present several years to a few months. Further, the engineered post translational tools will be applied to develop groundbreaking biotechnology applications enabled by orthogonal split-intein mediated protein ligation including manufacturing novel anti-reflection coating biomaterials and modular versatile multipecific antibodies at a later stage of this fellowship program. The successful outcome will lead to a transformative enabling technology for synthetic biology and open up a wide range of innovative protein-based biomanufacturing applications in the biotechnology and therapeutic sectors.