通过单分子廉价 FISH 与免疫荧光相结合对端粒酶 RNA 的成像。
Imaging of Telomerase RNA by Single-Molecule Inexpensive FISH Combined with Immunofluorescence.
机构信息
Department of Biochemistry and Molecular Medicine, Université de Montréal, Montréal, QC H3C 3J7, Canada.
Skirball Institute of Biomolecular Medicine, Cell Biology Department, NYU School of Medicine, New York, NY 10016, USA.
出版信息
STAR Protoc. 2020 Sep 10;1(2):100104. doi: 10.1016/j.xpro.2020.100104. eCollection 2020 Sep 18.
Fluorescent 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).
荧光原位杂交（FISH）检测端粒酶（hTR）的 RNA 部分与 50 个碱基探针检测 Cajal 体中积累的 hTR RNA。通过使用活细胞成像和单分子廉价 FISH，我们的已发表工作表明，只有一小部分 hTR 定位于 Cajal 体，而大多数 hTR 分子分布在核质中。本方案是 smiFISH 方法的应用指南，用于通过免疫荧光双重检测 hTR RNA、端粒或 Cajal 体。有关此方案使用和执行的完整详细信息，请参考 Laprade 等人（2020 年）。
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