Cytoplasmic DNA may arise upon DNA damage in the main nucleus where replication intermediates are cleaved during repair and released into the cytoplasm. Other sources of cytoplasmic DNA include the release of mitochondrial DNA (mtDNA) and micronuclear DNA. Such molecules have been shown to be recognized by cGAS, which facilitates the generation of cGAMP and subsequent downstream signaling by activation and phosphorylation of ER-associated STING. This triggers a phosphorylation cascade involving many factors, including TBK1 and various IKK family proteins. Similarly, DNA damage results in the release of RNA into the cytoplasm as a result of deregulated transcription, especially of repetitive elements including SINEs, as well as the release of mtRNA. These molecules are sensed by RIG-I and MDA5, depending on their length, and trigger immune signaling by engagement with mitochondrial membrane–associated MAVS. Upon activation, MAVS is polyubiquitinated by factors such as TRIM25 and engages in a phosphorylation cascade similar to that of activated STING. Ultimately, both of these result in the phosphorylation and nuclear translocation of IRF3 and NF-κB, which induces transcription of innate immune effector genes.