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Updated: Oct 6, 2025
High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay
Published on: June 1, 2014
Development of a single-step fluorogenic sirtuin assay and its applications for high-throughput screening.
Pan Wang1, Di Chen1, Jian-Xiong An1
1State Key Laboratory of Functions and Applications of Medicinal Plants, Engineering Research Center for the Development and Application of Ethnic Medicine and TCM (Ministry of Education), Guizhou Provincial Key Laboratory of Pharmaceutics, School of Pharmacy, Guizhou Medical University, Guiyang 550004, China. binhe@gmc.edu.cn.
Researchers developed novel fluorogenic probes to rapidly discover sirtuin (SIRT) modulators. These tools enable high-throughput screening for potential SIRT inhibitors, aiding in understanding SIRT roles in health and disease.
Area of Science:
- Biochemistry and Molecular Biology
- Chemical Biology
- Drug Discovery
Background:
- Sirtuins (SIRTs) are NAD+-dependent deacetylases crucial for biological functions.
- Dysregulation of SIRTs is implicated in various human diseases.
- A need exists for efficient methods to identify SIRT modulators.
Purpose of the Study:
- To design and synthesize novel fluorogenic probes for SIRT activity.
- To establish a high-throughput screening (HTS) method for SIRT modulators.
- To validate the utility of these probes in identifying SIRT inhibitors.
Main Methods:
- Design and synthesis of small molecules (4a-d) as fluorogenic probes.
- Probes utilize lysine substrates specific to SIRTs.
- Assay optimization, validation, kinetic studies, and HTS format application.
Main Results:
- Successful synthesis of four fluorogenic probes (4a-d).
- Demonstrated SIRT-catalyzed deacylation leading to fluorescence.
- Comprehensive characterization and validation of probes for HTS.
Conclusions:
- The developed fluorogenic probes are effective tools for SIRT modulation discovery.
- These probes facilitate rapid and accurate identification of SIRT inhibitors.
- The methodology supports efficient drug discovery for SIRT-related diseases.
