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High-resolution measurement of individual telomere lengths with Telo-seq

Nature Reviews Cancer volume 25, page 76 (2025)Cite this article

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Telomeres are the protective, repetitive DNA sequences at chromosome ends, and they are known to have a crucial role in ageing, development and diseases including cancer. Mammalian telomeres consist of 5′-TTAGGG-3′ repeats that terminate in a single-stranded 3′ overhang and shorten with each cell division. Existing methods for measuring telomere length, including terminal restriction fragment analysis using Southern blotting, next-generation sequencing and fluorescence in situ hybridization (FISH)-based methods, lack the nucleotide resolution, chromosomal annotation or key information about the entire telomere. By contrast, native long-read sequencing of full-length telomeres resolves the specific composition and length of all telomeres within a sample.

To measure the precise telomere length and sequence composition of all individual human chromosomes, we developed telomere sequencing (Telo-seq), a method to enrich and sequence all telomeres within a sample using Oxford Nanopore Technologies’ long-read sequencing. The workflow uses telorette (single-stranded oligonucleotide)-based adapters that anneal to the 3′ overhang, allowing sequencing through the long repetitive telomere and into the adjacent subtelomere region. The long-read approach overcomes the high sequence homology challenges associated with short-read technologies to capture enough unique subtelomeric sequences to distinguish between the different chromosome arms. In addition, sequencing native telomeres directly and without amplification with Oxford Nanopore Technologies’ devices retains the raw methylation signal. Thus, in combination with modification-sensitive basecallers, Telo-seq is able to analyse the subtelomeric methylation status.

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  1. Oxford Nanopore Technologies, San Francisco, CA, USA

    Carly Tyer

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  1. Carly Tyer
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Correspondence to Carly Tyer.

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C.T. is an employee of Oxford Nanopore Technologies, Inc. and is a stock or stock option holder. C.T. is named as an inventor on a patent application covering aspects of this work filed by Oxford Nanopore Technologies plc. Oxford Nanopore Technologies products are not intended for use for health assessment or to diagnose, treat, mitigate, cure or prevent any disease or condition.

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Tyer, C. High-resolution measurement of individual telomere lengths with Telo-seq. Nat Rev Cancer 25, 76 (2025). https://doi.org/10.1038/s41568-024-00767-7

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  • DOI: https://doi.org/10.1038/s41568-024-00767-7

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