Designing Artificial Cells towards a New Generation of Biosensors

Boyd, Margrethe; Kamat, Neha P.

doi:10.1016/j.tibtech.2020.12.002

Cited by 54 publications

(45 citation statements)

References 76 publications

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“…Moving forward, we can gain even finer control of membrane permeability by incorporating transmembrane proteins to enhance sensing capabilities and introduce more advanced sensing or responsive functions. These strategies could ultimately allow new functions for these types of sensors, including conjugation-based capture methods, deployment and transport of cell-free reactions, controlled sensor degradation, or enhanced sensor biocompatibility (18).…”

Section: Discussionmentioning

confidence: 99%

“…The copyright holder for this preprint this version posted March 2, 2022. ; https://doi.org/10.1101/2022.03.02.482665 doi: bioRxiv preprint Encapsulation enables tuning of the reaction environment on a molecular scale, enabling control of molecular interactions and addition of active membrane features to advance sensing capabilities, all while maintaining many of the tunable, advantageous features of cell-free systems (18).…”

Section: Introductionmentioning

confidence: 99%

“…Encapsulation enables tuning of the reaction environment on a molecular scale, enabling control of molecular interactions and addition of active membrane features to advance sensing capabilities, all while maintaining many of the tunable, advantageous features of cell-free systems (18).…”

Section: Introductionmentioning

confidence: 99%

“…By reconstituting purified cellular machinery in vitro , these systems enable use of natural microbial sensing mechanisms in a low-cost, distributable, and easily tunable platform. Despite these key advantages, removal from the cell also eliminates certain features of the cell’s native membrane barrier - such as reaction containment, protection from reaction inhibitors, and selective gating - all of which can add important functionality to cell-free biosensors (18).…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

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%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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

Boyd

Thavarajah

Lucks

et al. 2022

Preprint

Self Cite

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show abstract

“…50,51 This work, in particular, offers a platform for the development of lipid self-assemblies capable of sensing a chemical cue, offering a purely chemical perspective to cell-free biosensing. 52 In addition, our methodology enables investigation into the subcellular lipid environments that become targets of ONOO − during cellular nitrosative stress.…”

Section: Discussionmentioning

confidence: 99%

Direct Assessment of Nitrative Stress in Lipid Environments: Applications of a Designer Lipid-Based Biosensor for Peroxynitrite

Gutierrez

Erguven

Stone

et al. 2022

Preprint

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Peroxynitrite (ONOO−), a redox-active species generated under nitrosative stress conditions, can promote oxidative damage of lipids. Herein, we report the development and multifaceted utilization of a novel, activity-based phospholipid probe, PLP-ONOO–, for imaging lipid environments that are targeted by ONOO− in biomimetic and biological systems. Using PLP-ONOO– and the natural lipid 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, we built synthetic protocell membranes in the form of giant vesicles (GVs). These GVs responded to ONOO− by lighting up and displayed excellent selectivity against other redox-active species, introducing an unprecedented function to biomimetic membrane architectures. Live HeLa cells containing PLP-ONOO– were investigated under cellular nitrosative stress induced by either external administration of peroxynitrite or endogenous stimulation with interferon-γ / lipopolysaccharide / phorbol myristate acetate. Both conditions enhanced the coumarin fluorescence within cells, providing insight into intracellular lipid environments potentially prone to peroxynitrite-mediated oxidative damage. This work offers a chemical reactivity-dependent function for synthetic vesicles and opens a possibility for investigating nitrosative stress at subcellular levels.

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Artificial Receptor in Synthetic Cells Performs Transmembrane Activation of Proteolysis

Søgaard,

Løvschall,

Montasell

et al. 2024

Advanced Biology

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The design of artificial, synthetic cells is a fundamentally important and fast‐developing field of science. Of the diverse attributes of cellular life, artificial transmembrane signaling across the biomolecular barriers remains a high challenge with only a few documented successes. Herein, the study achieves signaling across lipid bilayers and connects an exofacial enzymatic receptor activation to an intracellular biochemical catalytic response using an artificial receptor. The mechanism of signal transduction for the artificial receptor relies on the triggered decomposition of a self‐immolative linker. Receptor activation ensues its head‐to‐tail decomposition and the release of a secondary messenger molecule into the internal volume of the synthetic cell. Transmembrane signaling is demonstrated in synthetic cells based on liposomes and mammalian cell‐sized giant unilamellar vesicles and illustrates receptor performance in cell mimics with a diverse size and composition of the lipid bilayer. In giant unilamellar vesicles, transmembrane signaling connects exofacial receptor activation with intracellular activation of proteolysis. Taken together, the results of this study take a step toward engineering receptor‐mediated, responsive behavior in synthetic cells.

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Designing Artificial Cells towards a New Generation of Biosensors

Cited by 54 publications

References 76 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

Direct Assessment of Nitrative Stress in Lipid Environments: Applications of a Designer Lipid-Based Biosensor for Peroxynitrite

Artificial Receptor in Synthetic Cells Performs Transmembrane Activation of Proteolysis

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