Synthetic biology – Engineering cell-based biomedical devices

Haellman, Viktor; Fussenegger, Martin

doi:10.1016/j.cobme.2017.09.010

Cited by 8 publications

(3 citation statements)

References 53 publications

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“…However, in order to introduce specific, adjustable, and scalable regulation, the information-processing pathways should preferably be designed de novo, as this minimizes the unwanted interactions with the cellular chassis and makes the pathways highly programmable for implementing designed cellular logic. Up to now, most designed cell circuits have been based on transcriptional regulation, drawing on the modular DNA recognition and transcriptional effector domains [7][8][9] . Transcriptional regulation-based cell logic is however inherently slower than protein interaction and modification-based systems.…”

Section: Introductionmentioning

confidence: 99%

Design of fast proteolysis-based signaling and logic circuits in mammalian cells

et al. 2018

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Cellular signal transduction is predominantly based on protein interactions and their posttranslational modifications, which enable a fast response to input signals. Due to difficulties in designing new unique protein-protein interactions, designed cellular logic has focused on transcriptional regulation; however, this has a substantially slower response requiring transcription and translation. Here, we present a de novo design of modular, scalable signaling pathways based on proteolysis and designed coiled-coils (CC) implemented in mammalian cells. A set of split proteases with highly specific orthogonal cleavage motifs was constructed and combined with strategically positioned cleavage sites and designed orthogonal CC dimerizing domains of tunable affinity for competitive displacement after proteolytic cleavage. This enabled implementation of Boolean logic functions and signaling cascades in mammalian cells. Designed split proteasecleavable orthogonal CC-based logic (SPOC logic) circuits enable response to chemical or biological signals within minutes rather than hours, useful for diverse medical and nonmedical applications.

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Section: Introductionmentioning

confidence: 99%

Design of fast proteolysis-based signaling and logic circuits in mammalian cells

et al. 2018

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“…Proteolytically modified effectors can irreversibly initiate signal transduction processes. Meanwhile, proteolytic regulation has been introduced into mammalian cells to control transcription factors and protein interactions [ 211 , 235 , 236 , 237 , 238 ] or even to develop cellular logic by cleaving transcription factors or bound proteins [ 239 , 240 , 241 ]. Since response rates of transcription factors are naturally slow, the application of proteolysis-based signaling systems to protein interactions and modifications is advantageous, as they occur within the range of minutes [ 211 ].…”

Section: Synthetic Biology Application To Crac Channels and Impact On...mentioning

confidence: 99%

Synthetic Biology Meets Ca2+ Release-Activated Ca2+ Channel-Dependent Immunomodulation

Bacsa,

Hopl,

Derler

2024

Cells

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Many essential biological processes are triggered by the proximity of molecules. Meanwhile, diverse approaches in synthetic biology, such as new biological parts or engineered cells, have opened up avenues to precisely control the proximity of molecules and eventually downstream signaling processes. This also applies to a main Ca2+ entry pathway into the cell, the so-called Ca2+ release-activated Ca2+ (CRAC) channel. CRAC channels are among other channels are essential in the immune response and are activated by receptor–ligand binding at the cell membrane. The latter initiates a signaling cascade within the cell, which finally triggers the coupling of the two key molecular components of the CRAC channel, namely the stromal interaction molecule, STIM, in the ER membrane and the plasma membrane Ca2+ ion channel, Orai. Ca2+ entry, established via STIM/Orai coupling, is essential for various immune cell functions, including cytokine release, proliferation, and cytotoxicity. In this review, we summarize the tools of synthetic biology that have been used so far to achieve precise control over the CRAC channel pathway and thus over downstream signaling events related to the immune response.

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“…Cellular complexity is governed by patterns of proteins that change in space and time. Recreating such patterns in cells or in a cell-free synthetic framework requires programming molecular interactions to regulate gene-expression reaction networks, − including recent effort toward a bioelectronic interface . The number of biological components that must be integrated scales up with the complexity of the network. , Biomolecular synthetic design would be simplified if molecular regulation of gene expression were replaced by external-field manipulation.…”

mentioning

confidence: 99%

Electric-Field Manipulation of a Compartmentalized Cell-Free Gene Expression Reaction

et al. 2018

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Direct electric-field manipulation of gene expression reactions would simplify the design of biochemical networks by replacing complex biomolecular interactions with push-button operations. Here, we applied a localized electric field gradient at megahertz frequency to manipulate a cell-free gene-expression reaction in a DNA compartment on a chip. We broke the spatial symmetry of a homogeneous reaction in the compartment by creating a trap for macromolecules in a region of maximal field intensity localized 50 μm from immobilized DNA. Free of biochemical regulation, we demonstrated protein synthesis oscillations by on/off switching of the electric field. In response to the field, ribosomes, RNA polymerases, and nascent RNA and proteins accumulated in the trap, and were then depleted from the DNA region where gene expression occurred. The resulting reduction in the rate of protein synthesis recovered back to steady-state when the field was off. The combination of electric field with compartmentalized cell-free gene expression reactions creates a simple, label-free approach for controlling biomolecules in space and time, opening possibilities for hybrid biological systems with a bioelectronic interface based on minimal biological parts design.

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Synthetic biology – Engineering cell-based biomedical devices

Cited by 8 publications

References 53 publications

Design of fast proteolysis-based signaling and logic circuits in mammalian cells

Design of fast proteolysis-based signaling and logic circuits in mammalian cells

Synthetic Biology Meets Ca2+ Release-Activated Ca2+ Channel-Dependent Immunomodulation

Electric-Field Manipulation of a Compartmentalized Cell-Free Gene Expression Reaction

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