title: In situ visualization of telomere elongation patterns in human cells.
abstract: The telomerase enzyme plays a critical role in human aging and cancer biology by maintaining telomere length and extending the proliferative lifespan of most stem cells and cancer cells. Despite the importance of this enzyme, our understanding of the mechanisms that regulate its activity and establish telomere length homeostasis in mammalian cells is incomplete, in part because the perfect repetitive nature of telomeric sequence hampers in situ detection of telomere elongation patterns. Here, we describe a novel assay using a mutant telomerase that adds a well-tolerated variant telomeric repeat sequence to telomere ends. By specifically detecting the addition of these variant repeats, we can directly visualize telomere elongation events in human cells. We validate this approach by in situ mapping of telomere elongation patterns within individual nuclei and across a population of cells.
authors: Department of Pathology, University of California, San Francisco, CA 94143, USA and Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94158, USA.
authors: Morgan E
authors: ME
authors: Diolaiti
authors: Beth A
authors: BA
authors: Cimini
authors: Robin
authors: R
authors: Kageyama
authors: Florie A
authors: FA
authors: Charles
authors: Bradley A
authors: BA
authors: Stohr
mesh_terms: D002460:Cell Line; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000235:genetics; D004247:DNA / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D009154:Mutation; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D016615:Telomere / Q000737:chemistry; D059505:Telomere Homeostasis
chemical_list: D004247:DNA; D012194:RNA-Directed DNA Polymerase
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