Comparison of telomere length measurement methods

• Published in: Philosophical transactions of the Royal Society of London. Series B. Biological sciences Vol. 373; no. 1741; p. 20160451
• Main Authors: Lai, Tsung-Po, Wright, Woodring E., Shay, Jerry W.
• Format: Journal Article
• Language: English
• Published: England The Royal Society 05.03.2018; The Royal Society Publishing
• Subjects: Animal tissues; Animals; Biomarkers; Cell fate; Cell proliferation; Cellular Senescence - genetics; Chromosomes; Deoxyribonucleic acid; DNA; DNA damage; Fluorescence; Fluorescence in situ hybridization; Humans; In Situ Hybridization, Fluorescence; Measurement methods; Polymerase chain reaction; Q-Fish; Q-Pcr; Real-Time Polymerase Chain Reaction; Reproducibility; Reproducibility of Results; Review; Senescence; Sensitivity and Specificity; Stela; Telomerase; Telomere - genetics; Telomere Homeostasis; Telomeres; Tesla; Trf; Validation Studies as Topic; Yeast; Q-PCR; STELA; TRF; TeSLA; Q-FISH
• ISSN: 0962-8436; 1471-2970
• DOI: 10.1098/rstb.2016.0451

The strengths and limitations of the major methods developed to measure telomere lengths (TLs) in cells and tissues are presented in this review. These include Q-PCR (Quantitative Polymerase Chain Reaction), TRF (Terminal Restriction Fragment) analysis, a variety of Q-FISH (Quantitative Fluorescence In Situ Hybridization) methods, STELA (Single TElomere Length Analysis) and TeSLA (Telomere Shortest Length Assay). For each method, we will cover information about validation studies, including reproducibility in independent laboratories, accuracy, reliability and sensitivity for measuring not only the average but also the shortest telomeres. There is substantial evidence that it is the shortest telomeres that trigger DNA damage responses leading to replicative senescence in mammals. However, the most commonly used TL measurement methods generally provide information on average or relative TL, but it is the shortest telomeres that leads to telomere dysfunction (identified by TIF, Telomere dysfunction Induced Foci) and limit cell proliferation in the absence of a telomere maintenance mechanism, such as telomerase. As the length of the shortest telomeres is a key biomarker determining cell fate and the onset of senescence, a new technique (TeSLA) that provides quantitative information about all the shortest telomeres will be highlighted. This article is part of the theme issue ‘Understanding diversity in telomere dynamics’.