title: Live-cell imaging of budding yeast telomerase RNA and TERRA.
abstract: In most eukaryotes, the ribonucleoprotein complex telomerase is responsible for maintaining telomere length. In recent years, single-cell microscopy techniques such as fluorescent in situ hybridization and live-cell imaging have been developed to image the RNA subunit of the telomerase holoenzyme. These techniques are now becoming important tools for the study of telomerase biogenesis, its association with telomeres and its regulation. Here, we present detailed protocols for live-cell imaging of the Saccharomyces cerevisiae telomerase RNA subunit, called TLC1, and also of the non-coding telomeric repeat-containing RNA TERRA. We describe the approach used for genomic integration of MS2 stem-loops in these transcripts, and provide information for optimal live-cell imaging of these non-coding RNAs.
authors.affiliation: Department of Biochemistry and Molecular Medicine, Université de Montréal, Montréal, Qc H3C 3J7, Canada.
authors.forename: Hadrien
authors.initials: H
authors.lastname: Laprade
authors.affiliation: Department of Biochemistry and Molecular Medicine, Université de Montréal, Montréal, Qc H3C 3J7, Canada.
authors.forename: Maxime
authors.initials: M
authors.lastname: Lalonde
authors.affiliation: Department of Biochemistry and Molecular Medicine, Université de Montréal, Montréal, Qc H3C 3J7, Canada.
authors.forename: David
authors.initials: D
authors.lastname: Guérit
authors.affiliation: Department of Biochemistry and Molecular Medicine, Université de Montréal, Montréal, Qc H3C 3J7, Canada. Electronic address: p.chartrand@umontreal.ca.
authors.forename: Pascal
authors.initials: P
authors.lastname: Chartrand
mesh_terms: D057054:Molecular Imaging / Q000379:methods*; D012313:RNA / Q000235:genetics*; D012331:RNA, Fungal / Q000235:genetics*; D022661:RNA, Untranslated / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D019098:Telomerase / Q000235:genetics*
chemical_list: D012331:RNA, Fungal; D022661:RNA, Untranslated; D029701:Saccharomyces cerevisiae Proteins; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase
keywords: MS2-GFP; TERRA; TLC1 RNA; Telomerase; Telomeres; Yeast