Isolation of endogenous cytosolic DNA from cultured cells

Shayla R Mosley; Kristi Baker

doi:10.1016/j.xpro.2022.101165

Isolation of endogenous cytosolic DNA from cultured cells

STAR Protoc. 2022 Feb 5;3(1):101165. doi: 10.1016/j.xpro.2022.101165. eCollection 2022 Mar 18.

Authors

Shayla R Mosley¹, Kristi Baker^{1

2}

Affiliations

¹ Department of Oncology, University of Alberta, Edmonton, AB T6G 2R3, Canada.
² Department of Medical Microbiology and Immunology, 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.

Keywords: Behavioral neuroscience; Biological sciences; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Cells, Cultured
Cytosol / metabolism
DNA* / genetics
Mitochondria* / genetics

Substances

DNA

Grants and funding

407882/CIHR/Canada