ArticleSTAR protocols2026
Protocol for condensate-based stabilization of gene circuit dynamics under growth-mediated dilution in E. coli.
Article in STAR protocols, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Growth-mediated dilution can destabilize synthetic gene circuits by reducing intracellular concentrations of key regulatory proteins. Here, we present a protocol that uses liquid-liquid phase separation to buffer dilution of transcription factors and stabilize synthetic gene circuits in E. coli. We describe steps for constructing phase-separating self-activation circuits, characterizing condensate material properties by fluorescence recovery after photobleaching, and imaging of condensate-promoter colocalization. We outline single-cell microscopy and population-level plate reader assays to quantify circuit activation, growth-coupled dilution dynamics, and memory retention. For complete details on the use and execution of this protocol, please refer to Zhang et al.
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