FRET-based reporter assesses lysosomal DNA-degradation ability in live cells

Area of Science:

• Cell Biology
• Molecular Biology
• Biochemistry

Background:

• Lysosomes are key organelles regulating cellular metabolism and homeostasis.
• Lysosomal dysfunction is linked to various diseases and adverse health outcomes.
• Current methods for assessing lysosomal function primarily focus on pH and protein degradation.

Purpose of the Study:

• To develop and validate a novel DNA-based reporter for evaluating lysosomal activity.
• To assess the DNA-degradation capacity of lysosomes as a functional readout.
• To investigate lysosomal function in disease models using this new reporter.

Main Methods:

• Development of a DNA-based reporter system.
• Utilizing fluorescence imaging to monitor lysosomal DNA-degradation ability.
• Application of the reporter in pharmacologically induced disease models (NP-A/B and NP-C).

Main Results:

• The DNA-based reporter successfully monitored lysosomal DNA-degradation ability.
• Dysregulated lysosomes and lysosomes in NP-A/B and NP-C models showed reduced DNA-degradation capacity.
• Pharmacological induction of disease models led to a significant decrease in lysosomal DNA-degradation ability.

Conclusions:

• The DNA-based reporter provides a novel tool for assessing lysosomal function.
• This method offers valuable insights into disease progression and therapeutic development.
• Monitoring lysosomal DNA-degradation ability is a promising approach for understanding lysosomal disorders.