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  1. The fusion of living bacteria and man-made materials represents a new frontier in medical and biosynthetic technology. However, the principles of bacterial signal processing inside synthetic materials with three-dimensional and fluctuating environments remain elusive. Here, we study bacterial growth in a three-dimensional hydrogel. We find that bacteria expressing an antibiotic resistance module can take advantage of ambient kinetic disturbances to improve growth while encapsulated. We show that these changes in bacterial growth are specific to disturbance frequency and hydrogel density. This remarkable specificity demonstrates that periodic disturbance frequency is a new input that engineers may leverage to control bacterial growth in synthetic materials. This research provides a systematic framework for understanding and controlling bacterial information processing in three-dimensional living materials. 
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  2. null (Ed.)
    Cell-free synthetic biology is a maturing field that aims to assemble biomolecular reactions outside cells for compelling applications in drug discovery, metabolic engineering, biomanufacturing, diagnostics, and education. Cell-free systems have several key features. They circumvent mechanisms that have evolved to facilitate species survival, bypass limitations on molecular transport across the cell wall, enable high-yielding and rapid synthesis of proteins without creating recombinant cells, and provide high tolerance towards toxic substrates or products. Here, we analyze ~750 published patents and ~2000 peer-reviewed manuscripts in the field of cell-free systems. Three hallmarks emerged. First, we found that both patent filings and manuscript publications per year are significantly increasing (five-fold and 1.5-fold over the last decade, respectively). Second, we observed that the innovation landscape has changed. Patent applications were dominated by Japan in the early 2000s before shifting to China and the USA in recent years. Finally, we discovered an increasing prevalence of biotechnology companies using cell-free systems. Our analysis has broad implications on the future development of cell-free synthetic biology for commercial and industrial applications. 
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  3. Abstract

    Synthetic biology has focused on engineering genetic modules that operate orthogonally from the host cells. A synthetic biological module, however, can be designed to reprogram the host proteome, which in turn enhances the function of the synthetic module. Here, we apply this holistic synthetic biology concept to the engineering of cell-free systems by exploiting the crosstalk between metabolic networks in cells, leading to a protein environment more favorable for protein synthesis. Specifically, we show that local modules expressing translation machinery can reprogram the bacterial proteome, changing the expression levels of more than 700 proteins. The resultant feedback generates a cell-free system that can synthesize fluorescent reporters, protein nanocages, and the gene-editing nuclease Cas9, with up to 5-fold higher expression level than classical cell-free systems. Our work demonstrates a holistic approach that integrates synthetic and systems biology concepts to achieve outcomes not possible by only local, orthogonal circuits.

     
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  4. The integration of synthetic biology and soft robotics can fundamentally advance sensory, diagnostic, and therapeutic functionality of bioinspired machines. However, such integration is currently impeded by the lack of soft-matter architectures that interface synthetic cells with electronics and actuators for controlled stimulation and response during robotic operation. Here, we synthesized a soft gripper that uses engineered bacteria for detecting chemicals in the environment, a flexible light-emitting diode (LED) circuit for converting biological to electronic signals, and soft pneu-net actuators for converting the electronic signals to movement of the gripper. We show that the hybrid bio-LED-actuator module enabled the gripper to detect chemical signals by applying pressure and releasing the contents of a chemical-infused hydrogel. The biohybrid gripper used chemical sensing and feedback to make actionable decisions during a pick-and-place operation. This work opens previously unidentified avenues in soft materials, synthetic biology, and integrated interfacial robotic systems. 
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