Gel-Free Tools for Quick and Simple Screening of Anti-Topoisomerase 1 Compounds

Area of Science:

• Biochemistry
• Molecular Biology
• Drug Discovery

Background:

• DNA topoisomerases are crucial enzymes regulating DNA topology, making them key targets for anti-cancer, anti-bacterial, and anti-parasitic drug development.
• Existing methods for assessing topoisomerase activity and compound inhibition are often time-consuming and lack laboratory adaptability.
• There is a growing need for efficient tools to investigate biomarkers and screen potential therapeutic compounds.

Purpose of the Study:

• To develop novel, rapid, and accessible screening methods for evaluating compounds that inhibit type 1 DNA topoisomerase activity.
• To create specific assays applicable to eukaryotic, viral, and bacterial topoisomerases for broad-spectrum drug discovery.

Main Methods:

• Development and implementation of rolling circle amplification (RCA)-based assays.
• Utilized RCA for screening small-molecule compounds against various type 1 topoisomerases, including human, *Leishmania donovani*, monkeypox virus, and *Mycobacterium smegmatis* topoisomerase 1.

Main Results:

• The developed RCA-based methods provide sensitive and directly quantitative readouts for topoisomerase inhibition.
• These assays are fast and easily adaptable, overcoming limitations of current screening techniques.
• Demonstrated successful application across diverse topoisomerase targets.

Conclusions:

• The novel RCA-based tools facilitate rapid drug screening and diagnostics for topoisomerase-targeted therapies.
• These methods can accelerate the development of new treatments for cancer and infectious diseases.
• Pave the way for improved research and clinical diagnostic protocols.