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Title: Photocaged DNAzymes as a general method for sensing metal ions in living cells

DNAzymes, which are sequences of DNA with catalytic activity, have been demonstrated as a potential platform for sensing a wide range of metal ions. Despite their significant promise, cellular sensing using DNAzymes has however been difficult, mainly because of the “always-on” mode of first-generation DNAzyme sensors. In this paper, to overcome this limitation, a photoactivatable (or photocaged) DNAzyme was designed and synthesized, and its application in sensing Zn lI in living cells was demonstrated. In this design, the adenosine ribonucleotide at the scissile position of the 8–17 DNAzyme was replaced by 2'-O-nitrobenzyl adenosine, rendering the DNAzyme inactive and thus allowing its delivery into cells intact, protected from nonspecific degradation within cells. Irradiation at 365 nm restored DNAzyme activity, thus allowing the temporal control over the sensing activity of the DNAzyme for metal ions. Finally, the same strategy was also applied to the GR-5 DNAzyme for the detection of PbII, thus demonstrating the possible scope of the method.
Authors:
 [1] ;  [1] ;  [1] ;  [2] ;  [1] ;  [2] ;  [1]
  1. Univ. of Illinois at Urbana-Champaign, Urbana, IL (United States)
  2. Univ. of California, San Diego, La Jolla, CA (United States)
Publication Date:
Grant/Contract Number:
FG02-08ER64568
Type:
Accepted Manuscript
Journal Name:
Angewandte Chemie (International Edition)
Additional Journal Information:
Journal Name: Angewandte Chemie (International Edition); Journal Volume: 53; Journal Issue: 50; Journal ID: ISSN 1433-7851
Publisher:
Wiley
Research Org:
Univ. of Illinois at Urbana-Champaign, IL (United States)
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; biosensors; DNAzymes; fluorescent probes; photolabile protecting groups
OSTI Identifier:
1342732

Hwang, Kevin, Wu, Peiwen, Kim, Taejin, Lei, Lei, Tian, Shiliang, Wang, Yingxiao, and Lu, Yi. Photocaged DNAzymes as a general method for sensing metal ions in living cells. United States: N. p., Web. doi:10.1002/anie.201408333.
Hwang, Kevin, Wu, Peiwen, Kim, Taejin, Lei, Lei, Tian, Shiliang, Wang, Yingxiao, & Lu, Yi. Photocaged DNAzymes as a general method for sensing metal ions in living cells. United States. doi:10.1002/anie.201408333.
Hwang, Kevin, Wu, Peiwen, Kim, Taejin, Lei, Lei, Tian, Shiliang, Wang, Yingxiao, and Lu, Yi. 2014. "Photocaged DNAzymes as a general method for sensing metal ions in living cells". United States. doi:10.1002/anie.201408333. https://www.osti.gov/servlets/purl/1342732.
@article{osti_1342732,
title = {Photocaged DNAzymes as a general method for sensing metal ions in living cells},
author = {Hwang, Kevin and Wu, Peiwen and Kim, Taejin and Lei, Lei and Tian, Shiliang and Wang, Yingxiao and Lu, Yi},
abstractNote = {DNAzymes, which are sequences of DNA with catalytic activity, have been demonstrated as a potential platform for sensing a wide range of metal ions. Despite their significant promise, cellular sensing using DNAzymes has however been difficult, mainly because of the “always-on” mode of first-generation DNAzyme sensors. In this paper, to overcome this limitation, a photoactivatable (or photocaged) DNAzyme was designed and synthesized, and its application in sensing ZnlI in living cells was demonstrated. In this design, the adenosine ribonucleotide at the scissile position of the 8–17 DNAzyme was replaced by 2'-O-nitrobenzyl adenosine, rendering the DNAzyme inactive and thus allowing its delivery into cells intact, protected from nonspecific degradation within cells. Irradiation at 365 nm restored DNAzyme activity, thus allowing the temporal control over the sensing activity of the DNAzyme for metal ions. Finally, the same strategy was also applied to the GR-5 DNAzyme for the detection of PbII, thus demonstrating the possible scope of the method.},
doi = {10.1002/anie.201408333},
journal = {Angewandte Chemie (International Edition)},
number = 50,
volume = 53,
place = {United States},
year = {2014},
month = {10}
}