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

利用单分子廉价 FISH 结合免疫荧光对端粒酶 RNA 进行成像

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作者:Emmanuelle Querido, Agnel Sfeir, Pascal Chartrand

Abstract

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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