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Red‐Shifted Fluorogenic Substrate for Detection of lac Z‐Positive Cells in Living Tissue with Single‐Cell Resolution

2018; Wiley; Volume: 57; Issue: 48 Linguagem: Inglês

10.1002/anie.201808670

1521-3773

Hiroki Ito, Yu Kawamata, Mako Kamiya, Kayoko Tsuda‐Sakurai, Shinji Tanaka, Tasuku Ueno, Toru Komatsu, Kenjiro Hanaoka, Shigeo Okabe, Masayuki Miura, Yasuteru Urano,

Biosensors and Analytical Detection

Abstract The Escherichia coli lac Z 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. Lac Z‐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.