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A Rapid and Versatile Assay for Ago2-Mediated Cleavage by Using Branched Rolling Circle Amplification
Marlen Hesse1, Christoph Arenz2
1Institut für Chemie, Humboldt-Universität zu Berlin, Brook-Taylor-Str. 2, 12489, Berlin, Germany.
Chembiochem : a European Journal of Chemical Biology
|December 18, 2015
Summary
We developed a novel assay for micro RNA (miRNA) target cleavage by Ago2. This cost-effective method uses branched rolling circle amplification for rapid detection, aiding in the discovery of miRNA inhibitors.
Area of Science:
- Molecular Biology
- Gene Regulation
- Biochemistry
Background:
- Micro RNA (miRNA) research is crucial for understanding gene regulation.
- Deregulation of miRNAs is linked to cellular dysfunction and various diseases.
- Investigating miRNA function is essential for disease research and therapeutic development.
Purpose of the Study:
- To develop a rapid, homogenous assay for Ago2-mediated target RNA cleavage.
- To enable the screening of small molecule inhibitors of miRNA function.
- To provide a cost-effective and practical method for miRNA research.
Main Methods:
- Utilized branched rolling circle amplification (BRCA) for detection.
- Developed a two-step assay involving Ago2-mediated cleavage and BRCA-based detection.
- Employed a label-free RNA substrate, avoiding artificial fluorescence labeling.
Main Results:
- Demonstrated a rapid and homogenous assay for Ago2-mediated RNA cleavage.
- The assay is capable of detecting small molecule inhibitors of miRNA function.
- The method is cost-effective, practicable, and compatible with standard qPCR machines.
Conclusions:
- The developed BRCA-based assay offers an efficient tool for studying miRNA target cleavage.
- This assay facilitates the screening of potential inhibitors for miRNA-related diseases.
- The label-free approach presents advantages for inhibitor screening and miRNA research.
