CRISPRi-Linked Multimodule Negative Feedback Loops to Address Winner-Take-All Resource Competition.

ABSTRACT: Cellular resource limitations create unintended interactions among synthetic gene circuit modules, compromising circuit modularity. This challenge is particularly pronounced in circuits with positive feedback, where uneven resource allocation can lead to Winner-Takes-All (WTA) behavior, favoring one module at the expense of others. In this study, we experimentally implemented a Negatively Competitive Regulatory (NCR) controller using CRISPR interference (CRISPRi) and evaluated its effectiveness in mitigating WTA behavior in two gene circuits: dual self-activation and cascading bistable switch. We chromosomally integrated a tunable dCas9 gene and designed module-specific gRNAs, with each module encoding its own gRNA to self-repress via competition for limited dCas9. This configuration intro

SUBMITTER: Rijal S 

PROVIDER: S-EPMC12641503 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature