Telomerase RNA Imaging in Budding Yeast and Human Cells by Fluorescent In Situ Hybridization

David Guérit; Maxime Lalonde; Pascal Chartrand

doi:10.1007/978-1-4939-7306-4_27

Telomerase RNA Imaging in Budding Yeast and Human Cells by Fluorescent In Situ Hybridization

Methods Mol Biol. 2018:1672:387-402. doi: 10.1007/978-1-4939-7306-4_27.

Authors

David Guérit¹, Maxime Lalonde¹, Pascal Chartrand²

Affiliations

¹ Department of Biochemistry and Molecular Medicine, Université de Montréal, CP 6128 Succ. Centre-ville, Montreal, H3C 3J7, QC, Canada.
² Department of Biochemistry and Molecular Medicine, Université de Montréal, CP 6128 Succ. Centre-ville, Montreal, H3C 3J7, QC, Canada. p.chartrand@umontreal.ca.

PMID: 29043638
DOI: 10.1007/978-1-4939-7306-4_27

Abstract

Telomerase, the enzyme that elongates telomeres in most eukaryotes, is a ribonucleoprotein complex composed of a reverse transcriptase catalytic subunit (TERT in human, Est2 in the budding yeast S. cerevisiae), regulatory factors and a noncoding RNA called hTERC (in human) or TLC1 (in budding yeast). Telomerase trafficking is a major process in the biogenesis and regulation of telomerase action at telomeres. Due to its higher signal-to-noise ratio, imaging of the telomerase RNA moiety is frequently used to determine telomerase intracellular localization. Here we describe how to image telomerase RNA in human and yeast cells using fluorescence in situ hybridization.

Keywords: Fluorescent in situ hybridization; RNA localization; TLC1 RNA; Telomerase; hTERC.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Humans
In Situ Hybridization, Fluorescence* / methods
Molecular Imaging* / methods
RNA / genetics*
Saccharomycetales / genetics*
Saccharomycetales / metabolism
Telomerase / genetics*

Substances

telomerase RNA
RNA
Telomerase

Grants and funding

MOP-89768/CIHR/Canada