In vitro selection of structure-switching, self-reporting aptamers
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SCOPUS
- Title
- In vitro selection of structure-switching, self-reporting aptamers
- Authors
- Oh, SS; Plakos, K; Lou, XH; Xiao, Y; Soh, HT
- Date Issued
- 2010-08-10
- Publisher
- National Academy of Sciences
- Abstract
- We describe an innovative selection approach to generate self-reporting aptamers (SRAs) capable of converting target-binding events into fluorescence readout without requiring additional modification, optimization, or the use of DNA helper strands. These aptamers contain a DNAzyme moiety that is initially maintained in an inactive conformation. Upon binding to their target, the aptamers undergo a structural switch that activates the DNAzyme, such that the binding event can be reported through significantly enhanced fluorescence produced by a specific stacking interaction between the active-conformation DNAzyme and a small molecule dye, N-methylmesoporphyrin IX. We demonstrate a purely in vitro selection-based approach for obtaining SRAs that function in both buffer and complex mixtures such as blood serum; after 15 rounds of selection with a structured DNA library, we were able to isolate SRAs that possess low nanomolar affinity and strong specificity for thrombin. Given ongoing progress in the engineering and characterization of functional DNA/RNA molecules, strategies such as ours have the potential to enable rapid, efficient, and economical isolation of nucleic acid molecules with diverse functionalities.
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/37728
- DOI
- 10.1073/PNAS.1009172107
- ISSN
- 0027-8424
- Article Type
- Article
- Citation
- PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 107, no. 32, page. 14053 - 14058, 2010-08-10
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