TY - JOUR
T1 - Red-Shifted Fluorogenic Substrate for Detection of lacZ-Positive Cells in Living Tissue with Single-Cell Resolution
AU - Ito, Hiroki
AU - Kawamata, Yu
AU - Kamiya, Mako
AU - Tsuda-Sakurai, Kayoko
AU - Tanaka, Shinji
AU - Ueno, Tasuku
AU - Komatsu, Toru
AU - Hanaoka, Kenjiro
AU - Okabe, Shigeo
AU - Miura, Masayuki
AU - Urano, Yasuteru
N1 - Funding Information:
This research was supported in part by AMED/CREST (JP18gm0710008 to Y.U.), by AMED (JP17gm0610004 and JP17gm5010001 to M.M.), by JST/PRESTO (JPMJPR14F8 to M.K.), by MEXT/JSPS KAKENHI; JP16H02606 and JP26111012 (to Y.U.), JP15H05951 “Resonance Bio” (to M.K.), 17H01387 (to S.O.), 16H06385 (to M.M.), by Brain/ MINDS (to Y.U.), by JSPS Core-to-Core Program, A. Advanced Research Networks, by a grant from Hoansha Foundation (to Y.U.), by Japan foundation for applied enzymology (to M.K.).
Publisher Copyright:
© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
PY - 2018/11/26
Y1 - 2018/11/26
N2 - 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.
AB - 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.
KW - fluorescent probes
KW - lacZ
KW - quinone methide
KW - single-cell resolution
KW - β-galactosidase
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U2 - 10.1002/anie.201808670
DO - 10.1002/anie.201808670
M3 - Article
C2 - 30255610
AN - SCOPUS:85055930312
SN - 1433-7851
VL - 57
SP - 15702
EP - 15706
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
IS - 48
ER -