Bioelectronic control of a microbial community using surface-assembled electrogenetic cells to route signals

Terrell, Jessica L.; Tschirhart, Tanya; Jahnke, Justin P.; Stephens, Kristina; Liu, Yi; Dong, Hong; Hurley, Margaret M.; Pozo, Maria; McKay, Ryan; Tsao, Chen Yu; Wu, Hsuan‐Chen; Vora, Gary J.; Payne, Gregory F.; Stratis‐Cullum, Dimitra N.; Bentley, William E.

doi:10.1038/s41565-021-00878-4

Cited by 72 publications

(102 citation statements)

References 56 publications

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“…The relay population was designed to detect hydrogen peroxide and respond by synthesizing AI-1. Plasmid pOxyRS-LasI ( Terrell et al, 2021 ) contains lasI , which synthesizes AI-1 under the hydrogen peroxide-sensitive oxyS promoter ( Fig. 2 a) and is adapted from a vector designed for peroxide induced gene expression ( Rubens et al, 2016 ).…”

Section: Resultsmentioning

confidence: 99%

“…2 a) and is adapted from a vector designed for peroxide induced gene expression ( Rubens et al, 2016 ). We previously modified lasI to incorporate a degradation tag ( Andersen et al, 1998 ) at the C-terminus to limit AI-1 production after the hydrogen peroxide has been consumed ( Terrell et al, 2021 ). In Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Bhokisham et al, used a similar system for electronic control of CRISPR components in spatially-controlled environments ( Bhokisham et al, 2020 ). More recently, Terrell et al generated hydrogen peroxide at an electrode surface to regulate a gene of interest from the oxyS promoter ( Terrell et al, 2021 ). That is, the OxyRS regulon is sensitive to hydrogen peroxide ( Virgile et al, 2018 ; Rubens et al, 2016 ), which in turn can be generated at a gold electrode surface through reduction of oxygen ( Qiang et al, 2002 ; Sanchez-Sanchez and Bard, 2009 ).…”

Section: Introductionmentioning

confidence: 99%

“…The second population that responds to the relayed signal and, in effect, controls the population profile, is referred to as the “controller” cells; they detect AI-1 (the signal relayed from the electrode) and grow faster in response. The relay cells, which detect hydrogen peroxide, are based on our previous work with cells that are adhered to a gold electrode ( Terrell et al, 2021 ). Here, we altered the design by showing that these cells need not be adhered to a gold electrode but can translate hydrogen peroxide signals while in the bulk media.…”

Section: Introductionmentioning

confidence: 99%

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Electronic signals are electrogenetically relayed to control cell growth and co-culture composition

Stephens

Zakaria

VanArsdale

et al. 2021

Metabolic Engineering Communications

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There is much to be gained by enabling electronic interrogation and control of biological function. While the benefits of bioelectronics that rely on potential-driven ionic flows are well known (electrocardiograms, defibrillators, neural prostheses, etc) there are relatively few advances targeting nonionic molecular networks, including genetic circuits. Redox activities combine connectivity to electronics with the potential for specific genetic control in cells. Here, electrode-generated hydrogen peroxide is used to actuate an electrogenetic “relay” cell population, which interprets the redox cue and synthesizes a bacterial signaling molecule (quorum sensing autoinducer AI-1) that, in turn, signals increased growth rate in a second population. The dramatically increased growth rate of the second population is enabled by expression of a phosphotransferase system protein, HPr, which is important for glucose transport. The potential to electronically modulate cell growth via direct genetic control will enable new opportunities in the treatment of disease and manufacture of biological therapeutics and other molecules.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Electronic signals are electrogenetically relayed to control cell growth and co-culture composition

Stephens

Zakaria

VanArsdale

et al. 2021

Metabolic Engineering Communications

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“…For example, a pentatyrosine protag was shown to allow tyrosinase-mediated covalent coupling of an IgG-binding protein G or a human glycoprotein, ApoH, with both polysaccharides (Wu et al, 2009) and silk fibroin from Bombyx mori (Wu et al, 2020); a polyglutamine tag facilitated the assembly of proteins onto both gelatin (Liu et al, 2015) and spider silk (Wu et al, 2017); and a polylysine tag was added to enzymes for covalent tethering onto engineered tobacco mosaic virus-derived virus like particles (Bhokisham et al, 2020). Other peptide tags of varied amino acid composition enable binding onto solid materials such as gold (Tamerler et al, 2006;Adams et al, 2015;Terrell et al, 2021), silver (Sedlak et al, 2012), silicon (Zhou et al, 2015), as well as various hydrophobic surfaces (Tanaka et al, 2006) through non-covalent interactions. Methodologies that allow protein attachment to various substrates have created new possibilities to construct devices with diverse functions introduced by the assembled proteins.…”

Section: Introductionmentioning

confidence: 99%

A Redox-Based Autoinduction Strategy to Facilitate Expression of 5xCys-Tagged Proteins for Electrobiofabrication

et al. 2021

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Biofabrication utilizes biological materials and biological means, or mimics thereof, for assembly. When interfaced with microelectronics, electrobiofabricated assemblies enable exquisite sensing and reporting capabilities. We recently demonstrated that thiolated polyethylene glycol (PEG-SH) could be oxidatively assembled into a thin disulfide crosslinked hydrogel at an electrode surface; with sufficient oxidation, extra sulfenic acid groups are made available for covalent, disulfide coupling to sulfhydryl groups of proteins or peptides. We intentionally introduced a polycysteine tag (5xCys-tag) consisting of five consecutive cysteine residues at the C-terminus of a Streptococcal protein G to enable its covalent coupling to an electroassembled PEG-SH film. We found, however, that its expression and purification from E. coli was difficult, owing to the extra cysteine residues. We developed a redox-based autoinduction methodology that greatly enhanced the yield, especially in the soluble fraction of E. coli extracts. The redox component involved the deletion of oxyRS, a global regulator of the oxidative stress response and the autoinduction component integrated a quorum sensing (QS) switch that keys the secreted QS autoinducer-2 to induction. Interestingly, both methods helped when independently employed and further, when used in combination (i.e., autodinduced oxyRS mutant) the results were best—we found the highest total yield and highest yield in the soluble fraction. We hypothesize that the production host was less prone to severe metabolic perturbations that might reduce yield or drive sequestration of the -tagged protein into inclusion bodies. We expect this methodology will be useful for the expression of many such Cys-tagged proteins, ultimately enabling a diverse array of functionalized devices.

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Living Synthelectronics: A New Era for Bioelectronics Powered by Synthetic Biology

Sun,

Yang,

et al. 2024

Advanced Materials

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Bioelectronics, which converges biology and electronics, has attracted great attention due to their vital applications in human–machine interfaces. While traditional bioelectronic devices utilize nonliving organic and/or inorganic materials to achieve flexibility and stretchability, a biological mismatch is often encountered because human tissues are characterized not only by softness and stretchability but also by biodynamic and adaptive properties. Recently, a notable paradigm shift has emerged in bioelectronics, where living cells, and even viruses, modified via gene editing within synthetic biology, are used as core components in a new hybrid electronics paradigm. These devices are defined as “living synthelectronics,” and they offer enhanced potential for interfacing with human tissues at informational and substance exchange levels. In this Perspective, the recent advances in living synthelectronics are summarized. First, opportunities brought to electronics by synthetic biology are briefly introduced. Then, strategic approaches to designing and making electronic devices using living cells/viruses as the building blocks, sensing components, or power sources are reviewed. Finally, the challenges faced by living synthelectronics are raised. It is believed that this paradigm shift will significantly contribute to the real integration of bioelectronics with human tissues.

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Bioelectronic control of a microbial community using surface-assembled electrogenetic cells to route signals

Cited by 72 publications

References 56 publications

Electronic signals are electrogenetically relayed to control cell growth and co-culture composition

Electronic signals are electrogenetically relayed to control cell growth and co-culture composition

A Redox-Based Autoinduction Strategy to Facilitate Expression of 5xCys-Tagged Proteins for Electrobiofabrication

Living Synthelectronics: A New Era for Bioelectronics Powered by Synthetic Biology

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