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Enzyme‐Mediated Endogenous and Bioorthogonal Control of a DNAzyme Fluorescent Sensor for Imaging Metal Ions in Living Cells
Author(s) -
Lin Yao,
Yang Zhenglin,
Lake Ryan J.,
Zheng Chengbin,
Lu Yi
Publication year - 2019
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201910343
Subject(s) - deoxyribozyme , bioorthogonal chemistry , fluorescence , biophysics , fluorophore , metal ions in aqueous solution , chemistry , cleavage (geology) , substrate (aquarium) , endogeny , dna , biochemistry , metal , combinatorial chemistry , biology , click chemistry , paleontology , ecology , physics , organic chemistry , quantum mechanics , fracture (geology)
Bioorthogonal control of metal‐ion sensors for imaging metal ions in living cells is important for understanding the distribution and fluctuation of metal ions. Reported here is the endogenous and bioorthogonal activation of a DNAzyme fluorescent sensor containing an 18‐base pair recognition site of a homing endonuclease (I‐ Sce I), which is found by chance only once in 7×10 10 bp of genomic sequences, and can thus form a near bioorthogonal pair with I‐ Sce I for DNAzyme activation with minimal effect on living cells. Once I‐ Sce I is expressed inside cells, it cleaves at the recognition site, allowing the DNAzyme to adopt its active conformation. The activated DNAzyme sensor is then able to specifically catalyze cleavage of a substrate strand in the presence of Mg 2+ to release the fluorophore‐labeled DNA fragment and produce a fluorescent turn‐on signal for Mg 2+ . Thus I‐ Sce I bioorthogonally activates the 10–23 DNAzyme for imaging of Mg 2+ in HeLa cells.