Bacterial two-component systems as sensors for synthetic biology applications

Lazar, John T.; Tabor, Jeffrey J.

doi:10.1016/j.coisb.2021.100398

Cited by 38 publications

(37 citation statements)

References 135 publications

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“…The diversity of existing cell-free sensors could ultimately lead to a new generation of encapsulated biosensors for a wide array of analytes. With the modularity of components in these systems, vesicle-based sensors could be engineered which use various membrane components, genetic circuits, and triggered responses to detect small molecules of interest (30, 56, 57). As focus shifts toward sensor application, these platforms could offer additional handles with which to tune sensor characteristics to advance the types of contexts in which cell-free sensing can operate, allowing for detection in environments like soil, ground water, or biological samples.…”

Section: Discussionmentioning

confidence: 99%

Robust and tunable performance of a cell-free biosensor encapsulated in lipid vesicles

Boyd

Thavarajah

Lucks

et al. 2022

Preprint

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Cell-free systems have enabled the development of genetically encoded biosensors to detect a range of environmental and biological targets. By encapsulating these systems in synthetic membranes, we can reintroduce features of the cell membrane, including molecular containment and selective permeability, which together could significantly enhance sensing capabilities. Here, we demonstrate robust and tunable performance of an encapsulated fluoride riboswitch inside of lipid vesicles. The riboswitch remains active upon encapsulation in lipid vesicles and responds to fluoride added to the surrounding solution. We find the sensitivity of the encapsulated sensor can be tuned by varying membrane composition. We then show that encapsulation protects the sensor from degradation by the sample and use two types of genetically encoded outputs to detect fluoride in real-world samples. This work establishes the feasibility of vesicle-encapsulated cell-free systems to detect environmentally relevant small molecules.

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Section: Discussionmentioning

confidence: 99%

Robust and tunable performance of a cell-free biosensor encapsulated in lipid vesicles

Boyd

Thavarajah

Lucks

et al. 2022

Preprint

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“…In bacteria, two-component signaling systems provide the best-known example of this mechanism. Two-component systems typically contain a transmembrane sensor protein and its associated cytosolic response regulator protein [ 57 , 58 ]. When a ligand comes into contact with the cell, it is recognized by the extracellular-facing ligand-binding domain of the sensor protein.…”

Section: Mechanistic Classes Of Biosensors Within Bacterial Cellsmentioning

confidence: 99%

Strategies for Improving Small-Molecule Biosensors in Bacteria

Miller

Bennett

2022

Biosensors

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In recent years, small-molecule biosensors have become increasingly important in synthetic biology and biochemistry, with numerous new applications continuing to be developed throughout the field. For many biosensors, however, their utility is hindered by poor functionality. Here, we review the known types of mechanisms of biosensors within bacterial cells, and the types of approaches for optimizing different biosensor functional parameters. Discussed approaches for improving biosensor functionality include methods of directly engineering biosensor genes, considerations for choosing genetic reporters, approaches for tuning gene expression, and strategies for incorporating additional genetic modules.

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“…temperature or pH), amino acids, succinate, and secondary metabolites [17][18][19][20] . Commonly, natural MRP encoded by CPAs is substantial and effective owing to their extreme importance for the survival of cells in fluctuating environments 21 . Therefore, simulating the natural MRP caused by environmental vibrations via CPA manipulation would be an effective strategy for reprogramming microbial metabolic networks.…”

Section: Mainmentioning

confidence: 99%

Harnessing cellular perception apparatus for smart metabolic reprogramming

Tan

Tao

2022

Preprint

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Metabolic reprogramming (MRP) is a fundamental approach in synthetic biology that involves redirecting metabolic flux and remodeling metabolic networks. However, only few approaches have been made in effective metabolic operations, especially at global level of metabolic networks. Naturally existing cellular perception apparatuses (CPAs), such as histidine kinases (HKs), are considered to sit on sensitive nodes of the metabolic network, which can trigger natural MRP upon perceiving environmental fluctuations. We develop a plateform for global MRP by natural environmental stimulation based on the combinational interference of CPAs. The plateform consists of a CRISPRi-mediated dual-gene combinational knockdown (CDCK) strategy and survivorship-based metabolic interaction analysis (SMIA). A total of 35 histidine kinase (HK) genes and 24 glycine metabolism genes were selected as targets to determine effectiveness of our approach for fast-growing chassis Vibrio FA2. Combined knockdown of several genes of HKs and glycine metabolism increased the glycine production. Other other hand, effects of CDCK on bacterial antibiotic resistance were assessed by targeting HKs. Many HKs were identified to be associated with antibiotic resistance in Vibrio FA2, of which combinational knockdown of two HK genes sasA_8 and 04288 reduced the ampicillin resistance. This MRP strategy is powerful and cost-effective, and can be considered as a smart strategy capable of operating a broad range of metabolic networks in microorganisms.

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Bacterial two-component systems as sensors for synthetic biology applications

Cited by 38 publications

References 135 publications

Robust and tunable performance of a cell-free biosensor encapsulated in lipid vesicles

Robust and tunable performance of a cell-free biosensor encapsulated in lipid vesicles

Strategies for Improving Small-Molecule Biosensors in Bacteria

Harnessing cellular perception apparatus for smart metabolic reprogramming

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