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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...

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Related Experiment Video

Updated: Jul 4, 2025

Droplet Digital TRAP ddTRAP: Adaptation of the Telomere Repeat Amplification Protocol to Droplet Digital Polymerase Chain Reaction

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Published on: May 3, 2019

Measuring telomerase activity using TRAP assays.

Gabriele Saretzki¹

¹Biosciences Institute, Newcastle University, Campus for Ageing and Vitality, Newcastle upon Tyne, United Kingdom.

Methods in Cell Biology

|February 1, 2024

Summary

The Telomerase Repeat Amplification Protocol (TRAP) measures telomerase activity, crucial for cancer diagnosis and therapy. This guide details TRAP methods, emphasizing controls and interpretation to avoid pitfalls in this complex technique.

Keywords:

Detection ELISA Measurement PCR TRAP assay Telomerase activity

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Last Updated: Jul 4, 2025

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Area of Science:

Molecular Biology
Biochemistry
Cancer Research

Background:

Telomerase, a reverse transcriptase (TERT/TERC), synthesizes telomeres at chromosome ends.
Telomerase activity is vital in germline, stem, and cancer cells.
Detecting telomerase activity (TA) is crucial for cancer diagnostics and therapy.

Purpose of the Study:

To describe various Telomerase Repeat Amplification Protocol (TRAP) methods for detecting telomerase activity (TA).
To provide guidance on performing and interpreting TRAP assays, including gel-based and ELISA methods.
To highlight the importance of controls and sample preparation for successful TRAP assay application.

Main Methods:

Detailed description of gel-based TRAP assays, including advantages and limitations.
Explanation of how to perform an Enzyme-Linked Immunosorbent Assay (ELISA)-based TRAP assay.
Brief mention of advanced TRAP modifications like real-time PCR, isothermal amplification, nanotechnology, and CRISPR/Cas-based methods.

Main Results:

TRAP assays enable the detection of telomerase activity in various cell types.
The chapter provides practical insights into assay execution and result interpretation.
Successful application of TRAP requires careful attention to controls and sample preparation.

Conclusions:

TRAP assays are indispensable tools for cancer clinical diagnosis and evaluating anti-cancer therapies targeting telomerase.
Understanding TRAP methodology, including potential pitfalls, is essential for reliable results.
This chapter serves as a practical guide to various TRAP techniques for researchers and clinicians.