Fluorogenic, Two-Photon-Triggered Photoclick Chemistry in Live Mammalian Cells

2013/10/29 by Zhipeng Yu, Tymish Y. Ohulchanskyy, Peng An +2 · 165 citations
Biochemistry, Genetics and Molecular Biology · Chemistry · Materials Science · #Advanced Fluorescence Microscopy Techniques #Biochemistry #Biophysics #Chemistry #Click Chemistry and Applications #Cycloaddition #Excitation #Femtosecond #Intracellular #Laser #Naphthalene #Organic chemistry #Photochemistry #Photochromic and Fluorescence Chemistry #Tetrazole #Two-photon excitation microscopy

paper · doi:10.1021/ja407867a

published in Journal of the American Chemical Society 135(45), 16766-16769 (American Chemical Society)

openalex publication_date 2013/10/29 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/01

Abstract

The tetrazole-based photoclick chemistry has provided a powerful tool to image proteins in live cells. To extend photoclick chemistry to living organisms with improved spatiotemporal control, here we report the design of naphthalene-based tetrazoles that can be efficiently activated by two-photon excitation with a 700 nm femtosecond pulsed laser. A water-soluble, cell-permeable naphthalene-based tetrazole was identified that reacts with acrylamide with the effective two-photon cross-section for the cycloaddition reaction determined to be 3.8 GM. Furthermore, the use of this naphthalene-tetrazole for real-time, spatially controlled imaging of microtubules in live mammalian cells via the fluorogenic, two-photon-triggered photoclick chemistry was demonstrated.

Citations

• Photoinducible Bioorthogonal Chemistry: A Spatiotemporally Controllable Tool to Visualize and Perturb Proteins in Live Cells

Cited by

• Light-Triggered Click Chemistry
• Photoclick Chemistry: A Bright Idea
• Examining Group Differences Between Suicidal Veterans Classified as Wish to Live, Ambivalent, or Wish to Die Using the Suicide Index Score
• Photoclick chemistry: a fluorogenic light-triggered in vivo ligation reaction. [europepmc]
• Design strategies for bioorthogonal smart probes. [europepmc]
• Development of bioorthogonal reactions and their applications in bioconjugation. [europepmc]
• Cyclopropenone-caged Sondheimer diyne (dibenzo[a,e]cyclooctadiyne): a photoactivatable linchpin for efficient SPAAC crosslinking. [europepmc]
• Light-Activated Staudinger-Bertozzi Ligation within Living Animals. [europepmc]
• Rapid Bioorthogonal Chemistry Turn-on through Enzymatic or Long Wavelength Photocatalytic Activation of Tetrazine Ligation. [europepmc]
• Novel Strategy for Preparing Dual-Modality Optical/PET Imaging Probes via Photo-Click Chemistry. [europepmc]
• Photo-Triggered Click Chemistry for Biological Applications. [europepmc]
• Sterically Shielded, Stabilized Nitrile Imine for Rapid Bioorthogonal Protein Labeling in Live Cells. [europepmc]
• Discovery of new mutually orthogonal bioorthogonal cycloaddition pairs through computational screening. [europepmc]
• Sterically shielded tetrazoles for a fluorogenic photoclick reaction: tuning cycloaddition rate and product fluorescence. [europepmc]
• An Extended Approach for the Development of Fluorogenic trans-Cyclooctene-Tetrazine Cycloadditions. [europepmc]
• Fitness Factors for Bioorthogonal Chemical Probes. [europepmc]
• Light-Triggered Click Chemistry. [europepmc]
• Light-induced primary amines and o-nitrobenzyl alcohols cyclization as a versatile photoclick reaction for modular conjugation. [europepmc]
• Oxidation-Induced "One-Pot" Click Chemistry. [europepmc]
• Photoclick Chemistry: A Bright Idea. [europepmc]
• Reversible Photoswitchable Inhibitors Generate Ultrasensitivity in Out-of-Equilibrium Enzymatic Reactions. [europepmc]
• Photo-controllable bioorthogonal chemistry for spatiotemporal control of bio-targets in living systems. [europepmc]
• A programmable chemical switch based on triggerable Michael acceptors. [europepmc]
• Enabling In Vivo Photocatalytic Activation of Rapid Bioorthogonal Chemistry by Repurposing Silicon-Rhodamine Fluorophores as Cytocompatible Far-Red Photocatalysts. [europepmc]
• Developing bioorthogonal probes to span a spectrum of reactivities. [europepmc]
• Bioorthogonal chemistry. [europepmc]
• Direct ring-strain loading for visible-light accelerated bioorthogonal ligation via diarylsydnone-dibenzo[b,f ][1,4,5]thiadiazepine photo-click reactions. [europepmc]
• Light-based fabrication and 4D customization of hydrogel biomaterials. [europepmc]

Related

• Photoinducible Bioorthogonal Chemistry: A Spatiotemporally Controllable Tool to Visualize and Perturb Proteins in Live Cells
• Targeted Covalent Photoswitch for Two-Photon Control of Endogenous Receptors
• Photo‐induced and Rapid Labeling of Tetrazine‐Bearing Proteins via Cyclopropenone‐Caged Bicyclononynes
• Multiphoton Activation of Photo-Strain-Promoted Azide Alkyne Cycloaddition “Click” Reagents Enables in Situ Labeling with Submicrometer Resolution
• Photoactivatable fluorophores for single-molecule localization microscopy of live cells