Red-Shifted Fluorogenic Substrate for Detection of lacZ-Positive Cells in Living Tissue with Single-Cell Resolution

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

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)15702-15706
Number of pages5
JournalAngewandte Chemie - International Edition
Volume57
Issue number48
DOIs
Publication statusPublished - 2018 Nov 26
Externally publishedYes

Keywords

  • fluorescent probes
  • lacZ
  • quinone methide
  • single-cell resolution
  • β-galactosidase

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)

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