GEN-Click: Genetically Encodable Click Reactions for Spatially Restricted Metabolite Labeling

ACS Cent Sci. 2023 Jul 25;9(8):1650-1657. doi: 10.1021/acscentsci.3c00511. eCollection 2023 Aug 23.


Chemical reactions for the in situ modification of biomolecules within living cells are under development. Among these reactions, bio-orthogonal reactions such as click chemistry using copper(I) and Staudinger ligation are widely used for specific biomolecule tracking in live systems. However, currently available live cell copper(I)-catalyzed azide/alkyne cycloaddition reactions are not designed in a spatially resolved manner. Therefore, we developed the “GEN-Click” system, which can target the copper(I)-catalyzed azide/alkyne cycloaddition reaction catalysts proximal to the protein of interest and can be genetically expressed in a live cell. The genetically controlled, spatially restricted, metal-catalyzed biorthogonal reaction can be used for proximity biotin labeling of various azido-bearing biomolecules (e.g., protein, phospholipid, oligosaccharides) in living cell systems. Using GEN-Click, we successfully detected local metabolite-transferring events at cell-cell contact sites.

PMID:37637744 | PMC:PMC10450880 | DOI:10.1021/acscentsci.3c00511


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