Project Summary: Administrative Supplement to NIGMS MIRA R35GM122561 Parent project title: Dynamics and evolution of synthetic and natural gene regulatory networks This Administrative Supplement is based on NOT-GM-24-021, “Notice of Special Interest (NOSI): Administrative Supplements for Equipment Purchases for Select NIGMS-Funded Awards”. It is a request for support for a “Jess” automated capillary-based western blot system by Bio-Techne, which allows for rapid, quantitative, highly reproducible, and multiplexed detection of up to 12 protein targets in a single sample without a need for genetical modification. These capabilities will be crucial for the parent project, which aims to learn how perturbed gene regulatory network dynamics and stochasticity affect single cells and thereby cell populations. To achieve this, we proposed using synthetic gene networks to generate specific gene expression patterns in space and time that serve as perturbation signals for natural gene networks, to study the subsequent effects on cell population behavior and evolution by computational modeling and experimental evolution. Unlike RNAs, which are relatively easy to quantify by sequencing, multi-protein quantification is still a challenge. Upon synthetic biological perturbation, the Jess system will uniquely unveil native regulatory network states at the protein level so that we can link them with cellular phenotypes. Overall, the Jess system will provide unique capabilities to illuminate at the protein level how regulatory networks enact control across scales of space and time in biology, from molecules to cells. This information will help us control adapting cell populations, which is relevant for understanding, predicting and possibly mitigating cancer and microbial drug resistance.