mtDNA-Specific Ultrasensitive Near-Infrared Fluorescent Probe Enables the Differentiation of Healthy and Apoptotic Cells

Yafu Wang, Huiyu Niu, Kui Wang, Ge Wang, Junwei Liu, Tony D. James, Hua Zhang

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Abstract

Mitochondrial DNA (mtDNA) as a class of important genetic material is easily damaged, which can result in a series of metabolic diseases, hereditary disease, and so on. mtDNA is an ultrasensitive indicator for the health of living cells due to the extremely short physiological response time of mtDNA toward damage (ca. 5.0 min). Therefore, the development of specific ultrasensitive fluorescent probes that can in real-time monitor mtDNA in vivo are of great value. With this research, we developed a near-infrared twisted intramolecular charge transfer (TICT) fluorescent probe YON. YON is a thread-like molecule with an A-π-D-π-A structure, based on the dicyanoisophorone fluorophore. The molecular design of YON enabled the specific binding with dsDNA (binding constant (K) = 8.5 × 105 M-1) within 1.3 min. And the appropriate water-oil amphiphilicity makes YON significantly accumulate in the mitochondria, enabling the specific binding to mtDNA. The fluorescence intensity at 640 nm of YON enhanced linearly with increasing concentrations of mtDNA. Dicyanoisophorone as the strong electron-withdrawing group that was introduced into both ends of the molecule resulted in YON being a classic quadrupole, so it could ultrasensitively detect trace mtDNA. The minimum detection limit was 71 ng/mL. Moreover, the large Stokes shift (λex = 435 nm, λem = 640 nm) makes YON suitable for "interference-free"imaging of mtDNA. Therefore, YON was used to monitor trace changes of mtDNA in living cells; more importantly, it could be used to evaluate the health of cells by monitoring microchanges of mtDNA, enabling the ultrasensitive evaluation of apoptosis.

Original languageEnglish
Pages (from-to)7510-7519
Number of pages10
JournalAnalytical Chemistry
Volume94
Issue number21
Early online date19 May 2022
DOIs
Publication statusPublished - 31 May 2022

Bibliographical note

Funding Information:
This work was supported by the National Natural Science Foundation of China (U21A20314, 22107089, 21722501). H.Z. wishes to thank the Zhongyuan high level talents special support plan-Science and Technology Innovation Leading Talents (204200510006). T.D.J. wishes to thank the Royal Society for a Wolfson Research Merit Award and the Open Research Fund of the School of Chemistry and Chemical Engineering Henan Normal University, for support (2020ZD01).

Funding Information:
This work was supported by the National Natural Science Foundation of China (U21A20314, 22107089, 21722501). H.Z. wishes to thank the Zhongyuan high level talents special support plan-Science and Technology Innovation Leading Talents (204200510006). T.D.J. wishes to thank the Royal Society for a Wolfson Research Merit Award and the Open Research Fund of the School of Chemistry and Chemical Engineering, Henan Normal University, for support (2020ZD01).

Publisher Copyright:
© 2022 American Chemical Society.

ASJC Scopus subject areas

  • Analytical Chemistry

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