Related Experiment Video
Updated: Sep 24, 2025
Preparation of Cytoplasmic and Nuclear Long RNAs from Primary and Cultured Cells
Published on: April 7, 2023
Isolation of endogenous cytosolic DNA from cultured cells
Shayla R Mosley1, Kristi Baker1,2
1Department of Oncology, University of Alberta, Edmonton, AB T6G 2R3, Canada.
Abstract:
DNA damage caused by genetic instability or extrinsic treatment can induce DNA leakage from the nucleus or mitochondria into the cytosol and activate innate and adaptive immunity. To enable characterization of these endogenous cytosolic DNAs and the mechanisms that produce them, we developed an approach for isolation of cytosolic DNA with no detectable mitochondrial contamination. Here we describe cytosolic compartment separation followed by DNA purification from colorectal cancer cells and illustrate how this may be expanded to other cell types.
Insights
DNA damage can release DNA into the cell
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- DNA damage from genetic instability or treatments can trigger immune responses.
- Cytosolic DNA, originating from the nucleus or mitochondria, activates innate and adaptive immunity.
Purpose of the Study:
- To develop a method for isolating cytosolic DNA without mitochondrial contamination.
- To enable the study of endogenous cytosolic DNAs and their production mechanisms.
Main Methods:
- Separation of the cytosolic compartment from colorectal cancer cells.
- DNA purification from the isolated cytosolic fraction.
Main Results:
- Successfully isolated cytosolic DNA with no detectable mitochondrial contamination.
- Demonstrated a method applicable to colorectal cancer cells.
Conclusions:
- The developed approach allows for the characterization of pure cytosolic DNA.
- This method can be expanded to other cell types for broader research applications.
