From scaffold to enzyme: Systematic engineering of the cofactor maturation scaffold NifEN reconstructs the P→M electron relay along with cofactor coordination and proton gating at the S2B site, conve....

Proteotoxic stress challenges multiple organelles, but how plants coordinate proteasome capacity with organellar function remains unclear. Langin et al. reveal that endoplasmic reticulum (ER)-associat...

Resource supply is generally considered a dominant driver of microbial diversity. Here, using a four-species bacterial consortium, we asked whether...

Photosynthetic eukaryotes have undergone evolutionary shifts from aquatic to terrestrial habitats, accompanied by changes in genome organization and gene regulation. Yet, the evolution of transfer RNA...

This study introduces genome–host association (GHA), leveraging natural epidemics to map the genetic landscape of host adaptation in the pathogen Zymoseptoria tritici. Applied to 832 wheat fungal stra...

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Investigating small-molecule dynamics within microbes is essential for comprehensive studies of microbial function. Both intra-organism and inter-organism small molecule dynamics play critical roles in microbial physiology, symbiosis, and disease. However, monitoring these dynamics remains highly challenging using most existing techniques. Fluorophore-based genetically encoded biosensors are powerful tools for tracking small-molecule dynamics in vivo and hold high potential for driving new discoveries. These biosensors are most commonly used in fluorescence imaging, often in combination with perfusion devices that allow precise control over environmental conditions. When integrated with advanced imaging techniques, this approach provides high-resolution, spatially and temporally resolved data, enabling insights into single-cell microbial responses. Despite their promise, implementing such biosensors remains technically challenging. Understanding the key steps is crucial for broader adoption. Here, we present a protocol designed to support the effective deployment of newly engineered biosensors into microbes for quantitative ratiometric fluorescence imaging under controlled conditions.

Investigating small-molecule dynamics within microbes is essential for comprehensive studies of microbial function. Both intra-organism and inter-organism small molecule dynamics play critical roles in microbial physiology, symbiosis, and disease. However, monitoring these dynamics remains highly challenging using most existing techniques. Fluorophore-based genetically encoded biosensors are powerful tools for tracking small-molecule dynamics in vivo and hold high potential for driving new discoveries. These biosensors are most commonly used in fluorescence imaging, often in combination with perfusion devices that allow precise control over environmental conditions. When integrated with advanced imaging techniques, this approach provides high-resolution, spatially and temporally resolved data, enabling insights into single-cell microbial responses. Despite their promise, implementing such biosensors remains technically challenging. Understanding the key steps is crucial for broader adoption. Here, we present a protocol designed to support the effective deployment of newly engineered biosensors into microbes for quantitative ratiometric fluorescence imaging under controlled conditions.