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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.

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Detecting Telomeric DNA Damage by Immuno-Telo FISH

  • 实验方案

  • 发表日期：

    2025年

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  • Marianna Paulis,
  • Ombretta Garbarino,
  • Francesca Faggioli

摘要

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.