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Telomeres and Telomerase

Telomeres and Telomerase

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 DNA.

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Updated: Jul 6, 2026

Telomere Length and Telomerase Activity; A Yin and Yang of Cell Senescence

Telomere Length and Telomerase Activity; A Yin and Yang of Cell Senescence

Published on: May 22, 2013

A sensitive direct human telomerase activity assay.

Scott B Cohen¹, Roger R Reddel

¹Cancer Research Unit, Children's Medical Research Institute, 214 Hawkesbury Road, Westmead, NSW 2145, Australia.

|April 1, 2008

Summary

This summary is machine-generated.

A new assay accurately measures human telomerase activity in cancer cells. This sensitive method uses immunoaffinity and DNA-substrate purification for reliable detection in research settings.

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Area of Science:

Biochemistry
Molecular Biology
Oncology

Background:

Telomerase, a reverse transcriptase, adds DNA repeats to chromosome ends.
Upregulated telomerase expression is a hallmark of over 85% of human cancers.
Sensitive detection of telomerase activity is crucial for cancer research.

Purpose of the Study:

To develop a direct primer-extension activity assay for human telomerase.
To achieve high sensitivity suitable for standard cell culture research.
To enable precise quantification of telomerase in cancer samples.

Main Methods:

Immunoaffinity purification of telomerase using specific antibodies and protein G-agarose beads.
Second affinity purification using neutravidin beads and a biotinylated telomeric DNA substrate.
Direct primer-extension assay on bead-immobilized telomerase and its DNA substrate.
Release and visualization of extension products via denaturing buffer and standard techniques.

Main Results:

The assay demonstrates sensitivity to approximately 10^6 telomerase-positive cells.
Over 90% of immunopurified telomerase is captured by the secondary affinity reagent.
The method provides highly enriched telomerase bound to its DNA substrate.
Telomerase extension products are successfully generated and detected.

Conclusions:

A sensitive and robust assay for human telomerase activity has been established.
The assay is suitable for cell culture-based research and cancer studies.
This method facilitates further investigation into telomerase function in cancer.