Development and application of novel BiFC probes for cell sorting based on epigenetic modification

Area of Science:

• Epigenetics
• Cancer Biology
• Biotechnology

Background:

• Cancer cell epigenetics change with disease progression and treatment.
• Understanding these changes at a single-cell level is crucial for tumor heterogeneity and progression studies.
• Live-cell compatible epigenome probes are needed for real-time analysis.

Purpose of the Study:

• To develop and validate novel bimolecular fluorescence complementation (BiFC) based epigenetic probes.
• To enable quantification and sorting of cells based on epigenetic modification content.
• To investigate the role of specific epigenetic marks (5mC and H3K9me3) in drug response.

Main Methods:

• Development of BiFC-based epigenetic probes for live-cell analysis.
• Application of flow cytometry for quantifying and sorting cells by epigenetic modifications.
• Assessment of sorted cell viability and response to chemotherapy drugs.

Main Results:

• Successfully developed and demonstrated BiFC probes for epigenetic profiling.
• Sorted cells remained viable and showed distinct responses to chemotherapy.
• Higher H3K9me3 levels correlated with Doxorubicin resistance in MCF7 cells.
• Higher 5mC levels correlated with increased responsiveness to Doxorubicin.

Conclusions:

• Novel split probes were successfully applied in flow cytometry for epigenetic analysis.
• The study elucidates the potential roles of 5-methylcytosine (5mC) and H3K9me3 in determining drug responses.
• This technology facilitates single-cell epigenetic studies for personalized cancer therapy.