Halo Tag: Difference between revisions
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== Introduction == | == Introduction == | ||
<StructureSection load='9HKF' size='340' side='right' caption='Crystal structure of psHaloTag labeled with JF635-HTL in the dark state' scene=''> | <StructureSection load='9HKF' size='340' side='right' caption='Crystal structure of psHaloTag labeled with JF635-HTL in the dark state' scene=''> | ||
The [Photoswitchable Halo Tag] (psHaloTag) is a unique chemigenetic system designed to provide reversible, light-controlled fluorescence tunable through both genetic and synthetic modifications, with promising applications for dynamic imaging in biological microscopy <ref>https://doi.org/10.1002/anie.202424955</ref>. psHaloTag addresses a major gap in the availability of reliable, reversible systems suitable for live-cell imaging. Many current systems are limited in tracking long-term, cyclical processes or in refreshing the pool of observable molecules. psHaloTag combines the genetically encoded HaloTag protein with the light-sensing sAsLOV2 domain. When illuminated with 450 nm light, psHaloTag undergoes a reversible change that activates a bound rhodamine dye ligand, resulting in a significant increase in fluorescence. This strong, multiple-cycle reporter overcomes the limitations of irreversible systems. It is an important tool for achieving precise spatiotemporal control in demanding applications such as live-cell Super-Resolution Microscopy (SMLM), where many established photosensitive probes cannot be reactivated. | The [Photoswitchable Halo Tag] (psHaloTag) is a unique chemigenetic system designed to provide reversible, light-controlled fluorescence tunable through both genetic and synthetic modifications, with promising applications for dynamic imaging in biological microscopy <ref> A Photoswitchable HaloTag for Spatiotemporal Control of Fluorescence in Living Cells https://doi.org/10.1002/anie.202424955</ref>. psHaloTag addresses a major gap in the availability of reliable, reversible systems suitable for live-cell imaging. Many current systems are limited in tracking long-term, cyclical processes or in refreshing the pool of observable molecules. psHaloTag combines the genetically encoded HaloTag protein with the light-sensing sAsLOV2 domain. When illuminated with 450 nm light, psHaloTag undergoes a reversible change that activates a bound rhodamine dye ligand, resulting in a significant increase in fluorescence. This strong, multiple-cycle reporter overcomes the limitations of irreversible systems. It is an important tool for achieving precise spatiotemporal control in demanding applications such as live-cell Super-Resolution Microscopy (SMLM), where many established photosensitive probes cannot be reactivated. | ||
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==== Mechanism ==== | ==== Mechanism ==== | ||
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The psHaloTag works as an allosteric | The psHaloTag works as an allosteric Photoswitch. | ||
*Protein Scaffold: An engineered HaloTag protein with the light-sensing sAsLOV2 domain. | *Protein Scaffold: An engineered HaloTag protein with the light-sensing sAsLOV2 domain. | ||
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====References==== | |||
<references/> | <references/> | ||
