Issue 3, 2016

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

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

Supplementary files

Article information

Article type
Edge Article
Submitted
21 Oct 2015
Accepted
01 Dec 2015
First published
01 Dec 2015
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2016,7, 1946-1953

Author version available

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

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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