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Issue 3, 2016
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Rational design of a quantitative, pH-insensitive, nucleic acid based fluorescent chloride reporter

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Abstract

Chloride plays a major role in cellular homeostasis by regulating the lumenal pH of intracellular organelles. We have described a pH-independent, fluorescent chloride reporter called Clensor that has successfully measured resting chloride in organelles of living cells. Here, we describe the rational design of Clensor. Clensor integrates a chloride sensitive fluorophore called 10,10′-bis[3-carboxypropyl]-9,9′-biacridinium dinitrate (BAC) with the programmability, modularity and targetability available to nucleic acid scaffolds. We show that simple conjugation of BAC to a DNA backbone fails to yield a viable chloride-sensitive reporter. Fluorescence intensity and lifetime investigations on a series of BAC-functionalized structural variants yielded molecular insights that guided the rational design and successful realization of the chloride sensitive fluorescent reporter, Clensor. This study provides some general design principles that would aid the realization of diverse ion-sensitive nucleic acid reporters based on the sensing strategy of Clensor.

Graphical abstract: Rational design of a quantitative, pH-insensitive, nucleic acid based fluorescent chloride reporter

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Publication details

The article was received on 21 Oct 2015, accepted on 01 Dec 2015 and first published on 01 Dec 2015


Article type: Edge Article
DOI: 10.1039/C5SC04002G
Citation: Chem. Sci., 2016,7, 1946-1953
  • Open access: Creative Commons BY license
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    Rational design of a quantitative, pH-insensitive, nucleic acid based fluorescent chloride reporter

    V. Prakash, S. Saha, K. Chakraborty and Y. Krishnan, Chem. Sci., 2016, 7, 1946
    DOI: 10.1039/C5SC04002G

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