Search bioRxiv⌕ Search

bioRxiv · 10.1101/2025.04.23.650302

Engineering microbial consortia for distributed signal processing

Abstract

A critical goal in biology is deducing input signals from measurable readouts. Genetic circuits have successfully been designed to respond to specific analytes and produce quantifiable outputs; however, multiplexed signal processing remains challenging. This limitation is partially due to crosstalk, or sensors non-specific responses to unintended signals. While strategies to achieve orthogonality are promising, they are time-intensive and context-dependent. Here, we introduce a new, generalizable approach that leverages microbial consortia to distribute sensory functions and computational methods to disentangle signals. Compartmentalizing sensor circuits within distinct populations simplifies experimental optimization by allowing individual populations to be exchanged without requiring genetic modifications. Our computational pipeline combines mechanistic modeling with machine learning to decode microbial communities unique temporal responses and predict multiple input concentrations. We demonstrated this platforms versatility in a variety of contexts: measuring signals with high crosstalk, detecting antibiotics with natural microbial communities, and quantifying chemicals in hospital sink wastewater. Our findings highlight how combining microbial engineering with computational strategies can produce robust, scalable biosensors for diverse applications.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Duncker, K. E., Shende, A. R., Shyti, I., Ruan, A., D'Cunha, R., Ma, H. R., Venugopal Lavanya, H., Liu, S., Gottel, N., Anderson, D. J., Gunsch, C. K., You, L.. 2025-04-24. Engineering microbial consortia for distributed signal processing. https://doi.org/10.1101/2025.04.23.650302

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Programmable interphyla communication for Staphylococcus aureus by transferability of homoserine lactone sensors between Bacillales bacteria

Staphylococcus aureus is a Gram-positive opportunistic pathogen of the order Bacillales and a frequent member of human-associated microbiomes. Despite its clinical relevance, engineering S. aureus and the communities it inhabits remains limited by a shortage of well-characterized genetic tools for precise control of gene expression and intercellular signaling. Biosensors for homoserine lactones (HSLs) are widely used in Gram-negative bacteria, as HSLs are small, passively diffusible signaling molecules for and their sensors are tightly regulated. Here, we show that synthetically designed HSL sensors using LuxR-family regulators are highly functional in S. aureus. Notably, synthetic designs previously built for the Gram-positive model Bacillus subtilis, a distantly related Bacillales, transferred functionally and establish repurposing the genetic toolbox as an effective strategy for importing these functions into S. aureus. Sensor performance correlated strongly between the two Bacillales, and activation in S. aureus reached 55-fold, exceeding the dynamic range of reported inducible systems. We further demonstrate tightly regulated, programmable interphyla communication between Gram-negative Escherichia coli and S. aureus via HSL signaling. These results provide a new toolset for engineering interbacterial communication and open a route to extending programmable HSL signaling to other tough-to-engineer bacteria.

synthetic biology↗

DIALECT: DNA-Programmable Synthetic Cells Communicate with Mammalian Cells through Synthetic Receptors

Synthetic cells can regulate mammalian signalling through the controlled release of diffusible protein ligands, but coupling programmable synthetic-cell senders to orthogonal, engineered mammalian receptors remains challenging. Here we present DIALECT (DNA-Instructed Assembly of Ligands for Engineered Cellular Transduction), a platform in which proteinosome-based synthetic cells translate sequence-defined DNA inputs into coiled-coil signalling ligand outputs for engineered mammalian receptors. DNA-ligand conjugates are localized within semipermeable proteinosomes through DNA hybridization and released by toehold-mediated DNA strand displacement (TMSD). The released ligands diffuse to neighbouring mammalian cells and activate cognate CC-GEMS receptors, inducing JAK/STAT3 signalling and reporter activation, with a higher response in cognate than non-cognate receiver cells under activation conditions. We further implement distributed Boolean AND logic using two independently programmed proteinosome populations that respond to distinct DNA inputs and release complementary ligand components. Simultaneous addition of both inputs produces substantially higher reporter activity than the respective single-input conditions, consistent with extracellular assembly of an active receptor agonist. DIALECT thus establishes a modular interface linking DNA-based molecular computation in synthetic cells to orthogonal signalling pathways in mammalian cells, providing a foundation for hybrid artificial-biological communication networks.

synthetic biology↗

Intracellular bio-synthesizing Gold Nanoparticles using bacterial systems and their applications in photothermal therapy (A green approach)

Background: Conventional cancer therapies are heavily constrained by systemic toxicity, off-target side effects, and poor deep-tissue penetration due to the highly heterogeneous tumor microenvironment (TME). Although gold nanoparticles (AuNPs) possess exceptional localized surface plasmon resonance (LSPR) properties ideal for photothermal therapy (PTT), their clinical translation is bottlenecked by inconsistent passive accumulation and chemical toxicity arising from conventional synthetic surfactants. Objective: This study developed a sustainable, biocompatible bio-hybrid platform utilizing the probiotic Escherichia coli Nissle 1917 (EcN) as an autonomous, tumor-homing living carrier integrated with AuNPs synthesized via an eco-friendly intracellular green chemistry approach. Methods: EcN cultures were prepared in Luria-Bertani (LB) medium and subsequently exposed to a 1 mM chloroauric acid (HAuCl4) metallic precursor. A facile, room-temperature green synthesis protocol was established by utilizing ascorbic acid (Vitamin C) as a non-toxic co-reductant to drive a seed-mediated crystallization onto intra-bacterial metallic seeds. The bio-hybrid system (EcN::AuNPs) was characterized via visual inspection, UV-Vis spectroscopy, and in vitro PTT assays under continuous-wave near-infrared (NIR) laser irradiation. Results: Visual confirmation showed an irreversible color shift from pale yellow to a deep dark-brownish hue, serving as a direct diagnostic indicator of successful Au3+ to Au0 reduction. UV-Vis spectroscopic analysis revealed a distinct, broadened surface plasmon resonance (SPR) band centered at 560nm, indicating the formation of stabilized, isotropic gold nanostructures (30-60nm) subjected to near-field electromagnetic coupling within the complex intracellular compartments (periplasm and cytoplasm). Biocompatibility assays demonstrated a significant collapse in cell viability from 10^8CFU/mL down to 10^3CFU/mL, driven by a combination of trivalent gold heavy-metal toxicity, mechanical disruption of the peptidoglycan wall from spatial confinement, and severe metabolic/oxidative stress. Nevertheless, the remaining surviving population maintained required active chemotactic capabilities, while the lysed bacterial matrix provided a natural protein capping layer that prevented nanoparticle aggregation. Under 5 minutes of continuous NIR laser irradiation, the EcN::AuNPs platform exhibited exceptional concentration-dependent hyperthermia kinetics, triggering a drastic thermal escalation from 20 C to an absolute peak of 75.4 C, vastly outperforming all baseline control groups (pure EcN, bare HAuCl4, and standalone ascorbic acid). Conclusion: The engineered EcN::AuNPs bio-hybrid platform represents a highly efficient and biosafe "bacteriobot" paradigm. By shifting the SPR footprint and leveraging the native bacterial cell envelope for colloidal stability, this system achieves an optimal equilibrium between active biological tumor targeting and rapid, localized photothermal ablation, offering a promising alternative for targeted oncological interventions.

synthetic biology↗