International Journal of Genetics and Genomics
Volume 7, Issue 3, September 2019, Pages: 60-68
Received: Jul. 1, 2019;
Accepted: Aug. 3, 2019;
Published: Aug. 23, 2019
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Fang Wang, Department of Obstetrics and Gynecology, Laboratory of Reproductive Medicine and NYU Fertility Center, NYU Langone Medical Center, New York, USAS
Leroy Robinson, Department of Obstetrics and Gynecology, Laboratory of Reproductive Medicine and NYU Fertility Center, NYU Langone Medical Center, New York, USAS
Yael Kramer, Department of Obstetrics and Gynecology, Laboratory of Reproductive Medicine and NYU Fertility Center, NYU Langone Medical Center, New York, USAS
Keri Kalmbach, Department of Obstetrics and Gynecology, Laboratory of Reproductive Medicine and NYU Fertility Center, NYU Langone Medical Center, New York, USAS
Paula Andrea Navarro, Department of Obstetrics and Gynecology, Laboratory of Reproductive Medicine and NYU Fertility Center, NYU Langone Medical Center, New York, USAS; Department of Obstetrics and Gynecology, USP Ribeirao Preto, Ribeirao Preto, Sao Paulo, Brazil
Ricardo Pimentel, Department of Obstetrics and Gynecology, Laboratory of Reproductive Medicine and NYU Fertility Center, NYU Langone Medical Center, New York, USAS
Xinghua Victor Pan, Department of Biochemistry and Molecular Biology, Southern Medical University, Ghuangzhou, China; Department of Genetics, Yale University School of Medicine, New Haven, USA
Sherman Weissman, Department of Genetics, Yale University School of Medicine, New Haven, USA
Lin Liu, Department of Obstetrics and Gynecology, Laboratory of Reproductive Medicine and NYU Fertility Center, NYU Langone Medical Center, New York, USAS; College of Life Sciences, Nankai University, Tianjin, China
David Keefe, Department of Obstetrics and Gynecology, Laboratory of Reproductive Medicine and NYU Fertility Center, NYU Langone Medical Center, New York, USAS
Increasing evidence demonstrates that shortest more than mean telomere length predicts telomere dysfunction and genomic instability in association with a number of conditions, including cell senescence, aging and tumorigenesis. We developed Universal Single Cell Single Telomere Length Analysis (USC-STELA), based on a PCR-amplification and southern blotting, to measure short telomeres in individual cells. The mean short telomere length measured in individual cells by USC-STELA correlates with that from bulk cells, measured by Universal STELA (U-STELA). The validation and reproducibility of USC-STELA was confirmed using different cell types with known telomere lengths, as well as by using paired sister-cells from human embryos and cultured cells. Interestingly, individual cells known to elongate telomeres via alternative lengthening of telomeres (ALT) have more short telomeres, yet longer mean telomere length than control cells. Moreover, individual senescent fibroblasts carry more short telomeres compared to human embryonic stem cells (hESCs), consistent with the notion that short telomeres contribute to cellular senescence. Additionally, we found a greater load of short telomeres in polar bodies than in matching oocytes, providing further insights into the accelerated polar body DNA degradation following extrusion from the oocyte. USC-STELA provides a new method to study telomere dysfunction in individual cells, with potential to improve our understanding of the role of telomere dynamics in cancer, developmental biology and reproductive medicine.
Paula Andrea Navarro,
Xinghua Victor Pan,
Measurement of Short Telomere Load in Individual Cells, International Journal of Genetics and Genomics.
Vol. 7, No. 3,
2019, pp. 60-68.
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