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Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis
Published on: July 8, 2025
Signaling aptamers created using fluorescent nucleotide analogues.
Evaldas Katilius1, Zivile Katiliene, Neal W Woodbury
1Department of Chemistry and Biochemistry and Center for BioOptical Nanotechnology, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA. EKatilius@asu.edu
Analytical Chemistry
|September 15, 2006
Summary
Researchers developed fluorescent aptamers for detecting proteins. This method uses nucleotide analogues that change fluorescence upon binding, enabling specific and sensitive detection of targets like thrombin.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Aptamers are nucleic acid-based tools for molecular recognition.
- Developing signaling aptamers with high sensitivity and specificity is crucial for diagnostics.
- Existing methods for aptamer-based detection can be limited by signal generation efficiency.
Purpose of the Study:
- To present a novel strategy for creating fluorescent signaling aptamers.
- To utilize the environment-sensitive fluorescence of nucleotide analogues for signal generation.
- To demonstrate the efficacy of this approach for detecting various protein targets.
Main Methods:
- Incorporation of fluorescent nucleotide analogues into aptamers at specific positions.
- Exploiting the dependence of fluorescence quantum yield on base stacking and conformational changes.
- Screening modified aptamer sequences and modeling secondary structures to identify optimal modification sites.
- Testing aptamer performance against human alpha-thrombin, immunoglobulin E, and platelet-derived growth factor B.
Main Results:
- Developed fluorescent signaling aptamers with binding-specific signal increases up to 30-fold.
- Identified key positions for nucleotide analogue modification through screening and modeling.
- Demonstrated successful application for detecting multiple protein targets.
- Showcased a robust method for generating fluorescence signals based on aptamer conformational changes.
Conclusions:
- The presented approach provides a versatile platform for engineering fluorescent aptamers.
- This method leverages the inherent properties of fluorescent nucleotide analogues for sensitive detection.
- The strategy is likely applicable to a broad range of protein-binding aptamers utilizing induced-fit mechanisms.
