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RNA and DNA aptamers in cytomics analysis
Henning Ulrich1, Antonio Henrique B Martins, João Bosco Pesquero
1Department of Biochemistry, Instituto de Química, Universidade de São Paulo, São Paulo, Brazil. henning@iq.usp.br
Summary
Systematic evolution of ligands by exponential enrichment (SELEX) generates aptamers for high-affinity protein binding. These aptamers are valuable tools for cytomics research and diagnostics, offering an alternative to traditional antibodies.
Area of Science:
- Biotechnology
- Molecular Biology
- Biochemistry
Background:
- Systematic evolution of ligands by exponential enrichment (SELEX) is a method for selecting DNA or RNA aptamers with high affinity and specificity for protein targets.
- Aptamers offer advantages over monoclonal antibodies, including in vitro generation and the ability to target non-immunogenic molecules.
Purpose of the Study:
- To demonstrate the in vitro selection of aptamers targeting the kinin B1 receptor.
- To explore the utility of aptamers in determining B1 receptor expression during inflammation.
Main Methods:
- In vitro selection using the SELEX technique to generate aptamers.
- Validation of aptamer specificity using Northern-Western blot assays.
- Application of aptamers for studying biological function and quantifying target concentrations in cytomics.
Main Results:
- Successfully selected anti-kinin B1 receptor aptamers through in vitro SELEX.
- Demonstrated specific recognition of the target protein by the selected aptamers in Northern-Western blot assays.
- Confirmed aptamer binding to the target protein when exposed on cell membranes.
Conclusions:
- Selected aptamers can be used to determine B1 receptor expression in inflammation.
- Aptamers are versatile tools for cytomics, enabling applications like fluorescence microscopy and flow cytometry.
- Future directions involve linking aptamers to fluorescent reporters for advanced cytomic analyses.