Imaging of Telomerase RNA by Single-Molecule Inexpensive FISH Combined with Immunofluorescence

Fluorescent in situ hybridization (FISH) on the RNA moiety of human telomerase (hTR) with 50-mer probes detects hTR RNA accumulated in Cajal bodies. Using both live-cell imaging and single-molecule inexpensive FISH, our published work revealed that only a fraction of hTR localizes to Cajal bodies, w...

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Detalles Bibliográficos
Autores principales: Querido, Emmanuelle, Sfeir, Agnel, Chartrand, Pascal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580239/
https://www.ncbi.nlm.nih.gov/pubmed/33111129
http://dx.doi.org/10.1016/j.xpro.2020.100104
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author Querido, Emmanuelle
Sfeir, Agnel
Chartrand, Pascal
author_facet Querido, Emmanuelle
Sfeir, Agnel
Chartrand, Pascal
author_sort Querido, Emmanuelle
collection PubMed
description Fluorescent in situ hybridization (FISH) on the RNA moiety of human telomerase (hTR) with 50-mer probes detects hTR RNA accumulated in Cajal bodies. Using both live-cell imaging and single-molecule inexpensive FISH, our published work revealed that only a fraction of hTR localizes to Cajal bodies, with the majority of hTR molecules distributed throughout the nucleoplasm. This protocol is an application guide to the smiFISH method for the dual detection of hTR RNA and telomeres or Cajal bodies by immunofluorescence. For complete details on the use and execution of this protocol, please refer to Laprade et al. (2020).
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spelling pubmed-75802392020-10-26 Imaging of Telomerase RNA by Single-Molecule Inexpensive FISH Combined with Immunofluorescence Querido, Emmanuelle Sfeir, Agnel Chartrand, Pascal STAR Protoc Protocol Fluorescent in situ hybridization (FISH) on the RNA moiety of human telomerase (hTR) with 50-mer probes detects hTR RNA accumulated in Cajal bodies. Using both live-cell imaging and single-molecule inexpensive FISH, our published work revealed that only a fraction of hTR localizes to Cajal bodies, with the majority of hTR molecules distributed throughout the nucleoplasm. This protocol is an application guide to the smiFISH method for the dual detection of hTR RNA and telomeres or Cajal bodies by immunofluorescence. For complete details on the use and execution of this protocol, please refer to Laprade et al. (2020). Elsevier 2020-09-10 /pmc/articles/PMC7580239/ /pubmed/33111129 http://dx.doi.org/10.1016/j.xpro.2020.100104 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Querido, Emmanuelle
Sfeir, Agnel
Chartrand, Pascal
Imaging of Telomerase RNA by Single-Molecule Inexpensive FISH Combined with Immunofluorescence
title Imaging of Telomerase RNA by Single-Molecule Inexpensive FISH Combined with Immunofluorescence
title_full Imaging of Telomerase RNA by Single-Molecule Inexpensive FISH Combined with Immunofluorescence
title_fullStr Imaging of Telomerase RNA by Single-Molecule Inexpensive FISH Combined with Immunofluorescence
title_full_unstemmed Imaging of Telomerase RNA by Single-Molecule Inexpensive FISH Combined with Immunofluorescence
title_short Imaging of Telomerase RNA by Single-Molecule Inexpensive FISH Combined with Immunofluorescence
title_sort imaging of telomerase rna by single-molecule inexpensive fish combined with immunofluorescence
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580239/
https://www.ncbi.nlm.nih.gov/pubmed/33111129
http://dx.doi.org/10.1016/j.xpro.2020.100104
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