Red-Shifted Fluorogenic Substrate for Detection of lacZ-Positive Cells in Living Tissue with Single-Cell Resolution.
Hiroki ItoYu KawamataMako KamiyaKayoko Tsuda-SakuraiShinji TanakaTasuku UenoToru KomatsuKenjiro HanaokaShigeo OkabeMasayuki MiuraYasuteru UranoPublished in: Angewandte Chemie (International ed. in English) (2018)
The Escherichia coli lacZ gene encoding β-galactosidase is a widely used reporter, but few synthetic substrates are available for detecting its activity with single-cell resolution in living samples. Our recently reported fluorogenic substrate SPiDER-βGal is suitable for this purpose, but its hydrolysis product shows green fluorescence emission, and a red-shifted analogue is therefore required for use in combination with green fluorescent protein (GFP) markers. Herein, we describe the development of a red-shifted fluorogenic substrate for β-galactosidase, SPiDER-Red-βGal, based on a silicon rhodol scaffold and a carboxylic group as the intramolecular nucleophile. LacZ-positive cells were successfully labeled with SPiDER-Red-βGal at single-cell resolution in living samples, which enabled us to visualize different cell types in combination with GFP markers.
Keyphrases
- single cell
- rna seq
- induced apoptosis
- escherichia coli
- single molecule
- high throughput
- cell cycle arrest
- amino acid
- computed tomography
- living cells
- genome wide
- stem cells
- structural basis
- label free
- binding protein
- mesenchymal stem cells
- pseudomonas aeruginosa
- cell proliferation
- candida albicans
- energy transfer
- genome wide identification