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Measuring Single-Cell Mitochondrial DNA Copy Number and Heteroplasmy Using Digital Droplet Polymerase Chain Reaction
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Detection and quantification of cytosolic DNA
Sheila Spada1, Takahiro Yamazaki1, Claire Vanpouille-Box1
1Department of Radiation Oncology, Weill Cornell Medicine, New York, NY, United States.
Methods in Enzymology
|November 16, 2019
Summary
Cytosolic DNA sensing, via the cGAS and STING pathway, is crucial for antitumor immunity. This chapter details methods to detect and quantify DNA in the cytosol, aiding cancer research.
Area of Science:
- Immunology
- Molecular Biology
- Oncology
Background:
- Cytosolic DNA sensing is a key mechanism initiating innate immune responses.
- The cyclic GMP-AMP synthase (cGAS) and stimulator of interferon genes (STING) pathway plays a critical role in this process.
- Understanding cytosolic DNA accumulation is vital for developing novel anticancer strategies.
Purpose of the Study:
- To present various techniques for detecting and quantifying cytosolic DNA.
- To provide researchers with essential methods for analyzing a critical readout in cancer immunology.
Main Methods:
- This chapter outlines diverse experimental approaches.
- Techniques focus on the detection and quantification of DNA within the cell's cytosol.
Main Results:
- The presented methods enable robust assessment of cytosolic DNA levels.
- Accurate quantification of cytosolic DNA is achievable through these techniques.
Conclusions:
- Effective detection and quantification of cytosolic DNA are essential for advancing antitumor immune strategies.
- The presented techniques offer valuable tools for researchers investigating the cGAS-STING pathway in cancer.

