Light-activated tetrazines enable precision live-cell bioorthogonal chemistry
Luping Liu1, Dongyang Zhang1, Mai Johnson1
1Department of Chemistry and Biochemistry, University of California, San Diego, CA, USA.
Nature Chemistry
|July 5, 2022
Summary
Researchers developed light-activated tetrazines for precise bioorthogonal labeling in living cells. This method enables spatiotemporal control for applications like cell membrane modification and photo-triggered drug delivery.
Area of Science:
- Bioconjugation Chemistry
- Chemical Biology
- Cellular Imaging
Background:
- Bioorthogonal reactions, like tetrazine cycloadditions, are vital for labeling biomolecules due to rapid kinetics.
- Achieving precise spatial and temporal control of these reactions in living mammalian cells remains a significant challenge.
- Existing tetrazine chemistries face limitations in stability and require methods for controlled activation.
Purpose of the Study:
- To develop a light-activated system for generating tetrazines in situ, enabling spatiotemporal control of bioorthogonal reactions.
- To overcome the stability issues of tetrazines and enable their early incorporation into biomolecules.
- To demonstrate the utility of this photoactivatable system for live-cell imaging and therapeutic applications.
Main Methods:
- Synthesis and characterization of photocaged dihydrotetrazines.
- Photouncaging strategy to generate reactive tetrazines upon light exposure.
- Application of the photoactivated tetrazines for bioorthogonal cycloaddition with trans-cyclooctenes in living cells.
- Labeling of cellular phospholipids and demonstration of photo-triggered prodrug release.
Main Results:
- Photocaged dihydrotetrazines are stable and can be activated by non-toxic light to form reactive tetrazines.
- The system allows for spatiotemporal control of bioorthogonal cycloadditions in living mammalian cells.
- Successful labeling of HeLa cell membranes with single-cell spatial resolution was achieved.
- Demonstrated proof-of-concept for photo-triggered therapy by coupling tetrazine ligation with prodrug release.
Conclusions:
- Light-activated tetrazine formation from photocaged dihydrotetrazines provides precise spatiotemporal control over bioorthogonal reactions in live cells.
- This approach enhances the stability and utility of tetrazine chemistry for biomolecule labeling and cellular imaging.
- The developed method holds promise for advanced applications in targeted drug delivery and photo-triggered therapies.
Related Concept Videos
01:19Protein Dynamics in Living Cells
2.2K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.2K
03:31Labeling DNA Probes
8.3K
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
8.3K