4.6 Article

Using the Intrinsic Fluorescence of DNA to Characterize Aptamer Binding

Journal

MOLECULES
Volume 27, Issue 22, Pages -

Publisher

MDPI
DOI: 10.3390/molecules27227809

Keywords

intrinsic fluorescence; DNA; aptamer; binding

Funding

  1. Natural Sciences and Engineering Research Council of Canada (NSERC)
  2. National Natural Science Foundation of China [31901776, 32072181]
  3. Agricultural Science and Technology Innovation Program [CAAS-ASTIP-2021-IFST-SN2021-05]
  4. China Scholarship Council (CSC) scholarship

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This study examined the possibility of using DNA intrinsic fluorescence to study aptamer binding. The results showed that DNA hybridization led to a decrease in fluorescence, while specific aptamers induced fluorescence increases. The length of the terminal stem had an impact on the fluorescence change for certain aptamers, while others failed to produce a fluorescence change.
The reliable, readily accessible and label-free measurement of aptamer binding remains a challenge in the field. Recent reports have shown large changes in the intrinsic fluorescence of DNA upon the formation of G-quadruplex and i-motif structures. In this work, we examined whether DNA intrinsic fluorescence can be used for studying aptamer binding. First, DNA hybridization resulted in a drop in the fluorescence, which was observed for A30/T30 and a 24-mer random DNA sequence. Next, a series of DNA aptamers were studied. Cortisol and Hg2+ induced fluorescence increases for their respective aptamers. For the cortisol aptamer, the length of the terminal stem needs to be short to produce a fluorescence change. However, caffeine and adenosine failed to produce a fluorescence change, regardless of the stem length. Overall, using the intrinsic fluorescence of DNA may be a reliable and accessible method to study a limited number of aptamers that can produce fluorescence changes.

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