Using confocal imaging and chemical crosslinking experiments, we confirmed that P2 can also report both on soluble oligomers and on insoluble aggregates of a POI fused with SNAP-tag in live cells. A new AggTag probe- P2, based on a SNAP-tag ligand bearing a green solvatochromic fluorophore-was synthesized for this purpose. In this study, we have expanded the AggTag method by using SNAP-tag technology to enable fluorogenic and biorthogonal detection of the aggregation of two different POIs simultaneously in live cells. ![]() However, the Halo-tag-based AggTag method only detects the aggregation of one specific POI at a time. We recently developed a fluorogenic method named aggregation tag (AggTag), and presented the AggTag probe P1, based on a Halo-tag ligand, to report on the aggregation of a protein of interest (POI) in live cells. However, numerous questions relating to protein aggregation remain unanswered due to the lack of available tools for visualization of these species in living cells. Protein aggregation involves the assembly of partially misfolded proteins into oligomeric and higher-order structures that have been associated with several neurodegenerative diseases.
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