2. Academic Validation
  3. Highly selective DNA aptamer sensor for intracellular detection of coenzyme A

Highly selective DNA aptamer sensor for intracellular detection of coenzyme A

  • Chem Sci. 2025 Mar 28;16(18):8023-8029. doi: 10.1039/d5sc00332f.
Yuan Ma 1 2 3,  Whitney Lewis 1 2,  Peng Yan 2 4,  Xiangli Shao 1 2,  Quanbing Mou 1 2 3,  Linggen Kong 5 6 7,  Weijie Guo 5 6 8,  Yi Lu 1 2 5 6 7
Affiliations

Affiliations

  • 1 Department of Chemistry, The University of Texas at Austin Austin Texas 78712 USA.
  • 2 Department of Chemistry, University of Illinois at Urbana-Champaign Urbana IL 61801 USA.
  • 3 Department of Chemistry, Rice University Houston TX 77005 USA.
  • 4 School of Health and Life Sciences, University of Health and Rehabilitation Sciences Qingdao Shandong 266113 P. R. China.
  • 5 Department of Molecular Biosciences, The University of Texas at Austin Austin Texas 78712 USA.
  • 6 Interdisciplinary Life Sciences Graduate Program, The University of Texas at Austin Austin Texas 78712 USA.
  • 7 Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign Urbana IL 61801 USA.
  • 8 Department of Biochemistry, University of Illinois at Urbana-Champaign Urbana IL 61801 USA.
PMID: 40206557 DOI: 10.1039/d5sc00332f
Abstract

Detecting Coenzyme A (CoA) in cells is vital for understanding its role in metabolism. DNA Aptamers, though widely used for monitoring many Other molecules, have not been effective for CoA detection, as previous attempts at obtaining DNA Aptamers for CoA using SELEX resulted in Aptamers that only recognize the adenine moiety of CoA. This "tyranny" of adenine dominating in SELEX has, therefore, hampered the SELEX of Aptamers specific for CoA. To meet this challenge, we employed a capture SELEX method by incorporating rigorous counter selections against adenine, adenosine, ATP, pantetheine, and pantothenic acid, resulting in a highly specific DNA aptamer for CoA over adenosine, ATP and Other related metabolites such as NADH, with a dissociation constant of 48.9 μM. This aptamer was then converted to a fluorescent sensor for CoA across pH 6.4-8.0. Confocal microscopy showed its ability to visualize CoA in living cells, with fluorescence changes observed upon manipulating CoA levels. This method broadens SELEX's application and presents a promising approach for studying and understanding CoA dynamics.

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