Tags

Type your tag names separated by a space and hit enter

Telomere Length Measurement by Molecular Combing.
Front Cell Dev Biol. 2020; 8:493.FC

Abstract

Telomeres are repetitive regions of DNA bound by specialized proteins at the termini of linear chromosomes that prevent the natural chromosome ends from being recognized as DNA double strand breaks. Telomeric DNA is gradually eroded with each round of cell division, resulting in the accumulation of critically short or dysfunctional telomeres that eventually trigger cellular senescence. Consequently, telomere length is indicative of the proliferative capacity of a cell. Multiple methods exist to measure telomere length and telomere content, but a simple and reliable technique to accurately measure individual telomere lengths is currently lacking. We have developed the Telomere length Combing Assay (TCA) to measure telomere length on stretched DNA fibers. We used TCA to measure telomere erosion in primary human fibroblasts, and to detect telomere lengthening in response to activation of telomere maintenance pathways. TCA was also used to accurately measure telomere length in healthy individuals, and to identify critically short telomeres in patients with telomere biology disorders. TCA is performed on isolated DNA, negating the need for cycling cells. TCA is amenable to semi-automated image analysis, and can be fully automated using the Genomic Vision molecular combing platform. This not only precludes sampling bias, but also provides the potential for high-throughput applications and clinical development. TCA is a simple and versatile technique to measure the distribution of individual telomere lengths in a cell population, offering improved accuracy, and more detailed biological insight for telomere length measurement applications.

Authors+Show Affiliations

Telomere Length Regulation Unit, Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW, Australia.Telomere Length Regulation Unit, Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW, Australia.Telomere Length Regulation Unit, Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW, Australia.Telomere Length Regulation Unit, Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW, Australia.Telomere Length Regulation Unit, Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW, Australia.Department of Hematology, Children's Hospital at Westmead, Sydney Children's Hospitals Network, Sydney, NSW, Australia.Department of Hematology, Children's Hospital at Westmead, Sydney Children's Hospitals Network, Sydney, NSW, Australia.Telomere Length Regulation Unit, Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW, Australia.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32612998

Citation

Kahl, Vivian F S., et al. "Telomere Length Measurement By Molecular Combing." Frontiers in Cell and Developmental Biology, vol. 8, 2020, p. 493.
Kahl VFS, Allen JAM, Nelson CB, et al. Telomere Length Measurement by Molecular Combing. Front Cell Dev Biol. 2020;8:493.
Kahl, V. F. S., Allen, J. A. M., Nelson, C. B., Sobinoff, A. P., Lee, M., Kilo, T., Vasireddy, R. S., & Pickett, H. A. (2020). Telomere Length Measurement by Molecular Combing. Frontiers in Cell and Developmental Biology, 8, 493. https://doi.org/10.3389/fcell.2020.00493
Kahl VFS, et al. Telomere Length Measurement By Molecular Combing. Front Cell Dev Biol. 2020;8:493. PubMed PMID: 32612998.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Telomere Length Measurement by Molecular Combing. AU - Kahl,Vivian F S, AU - Allen,Joshua A M, AU - Nelson,Christopher B, AU - Sobinoff,Alexander P, AU - Lee,Michael, AU - Kilo,Tatjana, AU - Vasireddy,Raja S, AU - Pickett,Hilda A, Y1 - 2020/06/16/ PY - 2020/02/14/received PY - 2020/05/25/accepted PY - 2020/7/3/entrez PY - 2020/7/3/pubmed PY - 2020/7/3/medline KW - Telomere length Combing Assay KW - senescence KW - telomerase KW - telomere KW - telomere length SP - 493 EP - 493 JF - Frontiers in cell and developmental biology JO - Front Cell Dev Biol VL - 8 N2 - Telomeres are repetitive regions of DNA bound by specialized proteins at the termini of linear chromosomes that prevent the natural chromosome ends from being recognized as DNA double strand breaks. Telomeric DNA is gradually eroded with each round of cell division, resulting in the accumulation of critically short or dysfunctional telomeres that eventually trigger cellular senescence. Consequently, telomere length is indicative of the proliferative capacity of a cell. Multiple methods exist to measure telomere length and telomere content, but a simple and reliable technique to accurately measure individual telomere lengths is currently lacking. We have developed the Telomere length Combing Assay (TCA) to measure telomere length on stretched DNA fibers. We used TCA to measure telomere erosion in primary human fibroblasts, and to detect telomere lengthening in response to activation of telomere maintenance pathways. TCA was also used to accurately measure telomere length in healthy individuals, and to identify critically short telomeres in patients with telomere biology disorders. TCA is performed on isolated DNA, negating the need for cycling cells. TCA is amenable to semi-automated image analysis, and can be fully automated using the Genomic Vision molecular combing platform. This not only precludes sampling bias, but also provides the potential for high-throughput applications and clinical development. TCA is a simple and versatile technique to measure the distribution of individual telomere lengths in a cell population, offering improved accuracy, and more detailed biological insight for telomere length measurement applications. SN - 2296-634X UR - https://www.unboundmedicine.com/medline/citation/32612998/Telomere_Length_Measurement_by_Molecular_Combing L2 - https://doi.org/10.3389/fcell.2020.00493 DB - PRIME DP - Unbound Medicine ER -
Try the Free App:
Prime PubMed app for iOS iPhone iPad
Prime PubMed app for Android
Prime PubMed is provided
free to individuals by:
Unbound Medicine.