A rapid SNAP-tag fluorogenic probe based on an environment-sensitive fluorophore for no-wash live cell imaging

Area of Science:

• Biochemistry
• Molecular Biology
• Chemical Biology

Background:

• Protein labeling with synthetic fluorophores often suffers from high background fluorescence.
• Extensive washing steps are typically required to remove unreacted fluorophores, complicating the process.

Purpose of the Study:

• To develop a novel fluorogenic probe for SNAP-tag proteins that overcomes the limitations of traditional labeling methods.
• To achieve highly sensitive and specific protein labeling with minimal background noise.

Main Methods:

• Development of a novel fluorogenic probe utilizing an environment-sensitive fluorophore.
• Application of the probe for labeling SNAP-tag proteins in vitro and in living cells.
• Assessment of fluorescence turn-on, selectivity, and labeling efficiency.

Main Results:

• The developed probe demonstrated a dramatic 280-fold fluorescence turn-on upon labeling with SNAP-tag.
• The probe exhibited high selectivity and rapid labeling kinetics with SNAP-tag proteins.
• Successful no-wash labeling of both intracellular and cell surface proteins was achieved in living cells.
• Specific visualization of protein localization was accomplished without extensive washing.

Conclusions:

• The novel fluorogenic probe offers a significant advancement for SNAP-tag protein labeling.
• Its dramatic fluorescence turn-on and no-wash capability simplify protein visualization in biological systems.
• This technology enables efficient and specific imaging of protein localization in living cells.