: FISH (fluorescence in situ hybridization) is one of the most reliable cytogenetic techniques for detecting changes in DNA integrity, including copy number gains and losses as well as telomeric erosion. Oncogene-induced senescence (OIS) is characterized by the persistence of DNA damage at telomeres. Immunofluorescence with γH2AX, a double-strand break marker, and PML, a tumor suppressor gene involved in OIS activation, can identify DNA lesions that activate the DNA repair machinery. Here, we describe a protocol by combining the two immuno-FISH approaches to detect dysfunctional telomeres in cell cultures, which can be used to monitor OIS under experimental conditions and potentially applied to ex vivo isolated senescent cells.
Detecting Telomeric DNA Damage by Immuno-Telo FISH
Paulis, MariannaPrimo
;Garbarino, Ombretta;Faggioli, Francesca
Ultimo
2025
Abstract
: FISH (fluorescence in situ hybridization) is one of the most reliable cytogenetic techniques for detecting changes in DNA integrity, including copy number gains and losses as well as telomeric erosion. Oncogene-induced senescence (OIS) is characterized by the persistence of DNA damage at telomeres. Immunofluorescence with γH2AX, a double-strand break marker, and PML, a tumor suppressor gene involved in OIS activation, can identify DNA lesions that activate the DNA repair machinery. Here, we describe a protocol by combining the two immuno-FISH approaches to detect dysfunctional telomeres in cell cultures, which can be used to monitor OIS under experimental conditions and potentially applied to ex vivo isolated senescent cells.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
