CRISPR-Stabilized Voronoi-like Domains in Synthetic Bacterial Populations

Abstract

Synthetic multicellular systems provide a controllable framework for studying how local interactions generate robust spatial organization in growing populations. Pattern formation is approached here as a programmable design problem, in which signaling, growth, and commitment must be coordinated to produce stable collective structure rather than treated as a secondary outcome. Voronoi-like tilings represent a useful class of organizations in which domains remain locally exclusive, collectively cover space, and often converge toward efficient hexagonal geometries, yet they have rarely been examined as emergent collective states governed by explicit local rules and tunable spatial control parameters. Here, a spatial-resolution-based framework is presented to generate stable Voronoi-like partitions in simulated bacterial colonies by combining quorum sensing, leader-follower role assignment, band-detection logic, and CRISPR-Cas9-mediated irreversible commitment. The system assigns local node identities, stabilizes spatial domains over time, and produces Voronoi-like partitions that emerge from distributed interactions rather than centralized control. The framework is implemented in the gro agent-based simulator and analyzed across parameter sweeps that vary leader-signal thresholds, node-size control, and signal propagation features. Robust spatial organization is found to depend on irreversible commitment, appropriate threshold separation, and controlled diffusion and degradation, revealing distinct robustness regimes that separate clean Voronoi-like partitions from deformed or mixed-identity patterns. These results position Voronoi-like spatial partitioning as a representative problem for programmable microbial active matter and support the use of synthetic gene circuits as minimal platforms for studying and exploiting spatial self-organization in microbial populations.