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Design and Synthesis of Caged Fluorescent Nucleotides and Application to Live‐cell RNA Imaging
Author(s) -
Ikeda Shuji,
Kubota Takeshi,
Wang Dan Ohtan,
Yanagisawa Hiroyuki,
Umemoto Tadashi,
Okamoto Akimitsu
Publication year - 2011
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201100523
Subject(s) - fluorescence , rna , nucleotide , nucleic acid , biophysics , chemistry , fluorescence lifetime imaging microscopy , photochemistry , microbiology and biotechnology , biochemistry , biology , optics , physics , gene
A binary photocontrolled nucleic acid probe that contains a nucleotide modified with one photolabile nitrobenzyl unit and two hybridization‐sensitive thiazole orange units has been designed for area‐specific fluorescence imaging of RNA in a cell. The synthesized probe emitted very weak fluorescence regardless of the presence of the complementary RNA, whereas it showed hybridization‐sensitive fluorescence emission at 532 nm after photoirradiation at 360 or 405 nm for uncaging. Fluorescence suppression of the caged probe was attributed to a decrease in the duplex‐formation ability. Caged fluorescent nucleotides with other emission wavelengths (622 and 724 nm) were also synthesized in this study; they were uncaged by 360 nm irradiation, and emitted fluorescence in the presence of the complementary RNA. Such probes were applied to area‐specific RNA imaging in a cell. Only probes in the defined irradiation area were activated by uncaging irradiation, and subnuclear mRNA diffusion in a living cell was monitored.