Diels–Alder Cycloaddition for Fluorophore Targeting to Specific Proteins inside Living Cells
Citations Over TimeTop 1% of 2011 papers
Abstract
The inverse-electron-demand Diels–Alder cycloaddition between trans-cyclooctenes and tetrazines is biocompatible and exceptionally fast. We utilized this chemistry for site-specific fluorescence labeling of proteins on the cell surface and inside living mammalian cells by a two-step protocol. Escherichia coli lipoic acid ligase site-specifically ligates a trans-cyclooctene derivative onto a protein of interest in the first step, followed by chemoselective derivatization with a tetrazine–fluorophore conjugate in the second step. On the cell surface, this labeling was fluorogenic and highly sensitive. Inside the cell, we achieved specific labeling of cytoskeletal proteins with green and red fluorophores. By incorporating the Diels–Alder cycloaddition, we have broadened the panel of fluorophores that can be targeted by lipoic acid ligase.
Related Papers
- → Tetrazine‐Isonitrile Bioorthogonal Fluorogenic Reactions Enable Multiplex Labeling and Wash‐Free Bioimaging of Live Cells(2024)38 cited
- → Overview of Syntheses and Molecular-Design Strategies for Tetrazine-Based Fluorogenic Probes(2021)49 cited
- → A tetrazine-fused aggregation induced emission luminogen for bioorthogonal fluorogenic bioprobe(2021)28 cited
- → Tetrazine‐Isonitrile Bioorthogonal Fluorogenic Reactions Enable Multiplex Labeling and Wash‐Free Bioimaging of Live Cells(2024)3 cited
- → Control of tetrazine bioorthogonal reactivity by naphthotube and phenyltetrazine host-guest pairs(2023)3 cited