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Related Concept Videos

Telomeres and Telomerase02:41

Telomeres and Telomerase

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In eukaryotic DNA replication, a single-stranded DNA fragment remains at the end of a chromosome after the removal of the final primer. This section of DNA cannot be replicated in the same manner as the rest of the strand because there is no 3’ end to which the newly synthesized DNA can attach. This non-replicated fragment results in gradual loss of the chromosomal DNA during each cell duplication. Additionally, it can induce a DNA damage response by enzymes that recognize single-stranded...
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Updated: May 22, 2025

Observation and Quantification of Telomere and Repetitive Sequences Using Fluorescence In Situ Hybridization FISH with PNA Probes in Caenorhabditis elegans
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Detecting Telomeric DNA Damage by Immuno-Telo FISH.

Marianna Paulis1,2, Ombretta Garbarino1, Francesca Faggioli3,4

  • 1Institute of Genetics and Biomedical Research, UoS of Milan, National Research Council, Milan, Italy.

Methods in Molecular Biology (Clifton, N.J.)
|March 14, 2025
PubMed
Summary

This study introduces a combined immuno-fluorescence in situ hybridization (FISH) method to detect DNA damage at telomeres, crucial for understanding oncogene-induced senescence (OIS). This technique helps identify dysfunctional telomeres and monitor OIS in cell cultures and senescent cells.

Keywords:
Immuno-teloFISHPMLTelomeresγH2AXOncogene-induced senescence

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Fluorescence In Situ Hybridization on DNA Halo Preparations to Reveal Whole Chromosomes, Telomeres and Gene Loci
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Area of Science:

  • Cytogenetics
  • Molecular Biology
  • Cellular senescence

Background:

  • Telomere erosion and DNA damage are hallmarks of oncogene-induced senescence (OIS).
  • Detecting DNA lesions that activate DNA repair is key to understanding OIS.
  • Existing methods may not fully capture telomere dysfunction in senescence.

Purpose of the Study:

  • To describe a novel protocol combining immuno-FISH to detect dysfunctional telomeres.
  • To provide a method for monitoring OIS in experimental settings.
  • To explore applications for analyzing ex vivo senescent cells.

Main Methods:

  • Utilizing fluorescence in situ hybridization (FISH) for DNA integrity assessment.
  • Employing immunofluorescence with γH2AX (DNA double-strand break marker) and PML (senescence marker).
  • Combining immuno-FISH to identify DNA lesions at telomeres.

Main Results:

  • The combined immuno-FISH protocol effectively detects dysfunctional telomeres.
  • The method identifies DNA damage associated with OIS activation.
  • The protocol is suitable for cell cultures and potentially ex vivo senescent cells.

Conclusions:

  • The developed immuno-FISH protocol is a reliable tool for studying telomere dysfunction in OIS.
  • This technique facilitates monitoring of OIS under experimental conditions.
  • The method has potential applications in analyzing senescent cells isolated from tissues.