Chemigenetic Encoding of Fluorescent Dyes Enables High-Fidelity and Wash-Free Imaging of Proteins in Live Cells.

ABSTRACT: Fluorescent dyes are indispensable chemical tools for protein labeling, yet their utility in live-cell imaging remains constrained by background fluorescence from off-target interactions. A chemigenetic strategy is presented that integrates synthetic dye chemistry with genetically encoded fluorescence to achieve high-fidelity, wash-free imaging of target proteins. By leveraging Förster resonance energy transfer (FRET) between fluorescent proteins (FPs) and Halo-tag dyes, a system is engineered where fluorescence emission depends on FPs donor excitation, ensuring only dyes bound to target protein are fluorescent, while nonspecifically bound dyes remain dark, enhancing the signal-to-noise ratio (SNR). This approach is implemented by fusing Halo-tag with FPs (sfGFP, mCherry) to create FRET pa

SUBMITTER: Zhou X 

PROVIDER: S-EPMC12407358 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature