DNA Repair Biosensor-Identified DNA Damage Activities of Endophyte Extracts from Garcinia cowa

Area of Science:

• Oncology
• Molecular Biology
• Biochemistry

Background:

• Chemotherapy increasingly targets cancer-specific mechanisms to minimize side effects.
• DNA damage and repair pathways are critical targets for novel cancer therapeutics.
• Targeting DNA repair offers a promising strategy for developing more effective and selective cancer treatments.

Purpose of the Study:

• To develop and validate DNA repair biosensor cells for screening natural compounds.
• To investigate the DNA repair activities induced by a natural compound extract.
• To identify potential therapeutic agents targeting DNA repair pathways in cancer.

Main Methods:

• Engineered cells with fluorescently tagged 53BP1 and RAD54L proteins to monitor DNA double-strand break (DSB) repair pathways (NHEJ and HR).
• Validated biosensor cell lines against wild-type cells for accurate DNA repair assessment.
• Tested a crude extract from Phyllosticta sp. (endophyte of Garcinia cowa) for its effect on DNA damage and repair.

Main Results:

• The engineered biosensor cells accurately reflected wild-type DNA repair properties.
• The Phyllosticta sp. extract induced DNA double-strand breaks (DSBs), evidenced by increased γH2AX.
• Evidence suggests the extract promotes DNA repair via the non-homologous end-joining (NHEJ) pathway, indicated by elevated 53BP1 focus formation.

Conclusions:

• DNA repair-based biosensors are effective tools for preliminary screening of natural compounds.
• The study identified a Phyllosticta sp. extract as a potential agent inducing DNA damage and activating NHEJ repair.
• This approach facilitates the discovery of novel therapeutic compounds for cancer treatment targeting DNA repair mechanisms.