Abstract
Telomeres play an important role in ensuring the integrity of the genome. Telomere shortening can lead to loss of genetic information and trigger DNA damage responses. Cultured mammalian cells have served as critical model systems for studying the function of telomere binding proteins and telomerase. Tremendous heterogeneity can be observed both between species and within a single cell population. Recent advances in genome editing (such as the development of the CRISPR/Cas9 platform) have further enabled researchers to carry out loss-of-function analysis of how disrupting key players in telomere maintenance affects telomere length regulation. Here we describe the steps to be carried out in order to analyze the average length of telomeres in CRISPR-engineered human knockout (KO) cells (TRF analysis).
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Acknowledgment
The authors thank Drs. Matthew O’Conner and Dong Yang for technical input and advice, and the generous support from the Dan L. Duncan Cancer Center and program grant P30CA125123.
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Xu, J., Songyang, Z., Liu, D., Kim, H. (2017). Analysis of Average Telomere Length in Human Telomeric Protein Knockout Cells Generated by CRISPR/Cas9. In: Songyang, Z. (eds) Telomeres and Telomerase. Methods in Molecular Biology, vol 1587. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6892-3_2
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DOI: https://doi.org/10.1007/978-1-4939-6892-3_2
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6891-6
Online ISBN: 978-1-4939-6892-3
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