Detection of a 7SL RNA-derived small non-coding RNA using Molecular Beacons in vitro and in cells

被引:2
|
作者
Weigert, Nina [1 ]
Schweiger, Anna-Lena [1 ]
Gross, Jonas [1 ]
Matthes, Marie [1 ]
Corbacioglu, Selim [1 ]
Sommer, Gunhild [1 ]
Heise, Tilman [1 ]
机构
[1] Univ Hosp Regensburg, Dept Pediat Hematol Oncol & Stem Cell Transplantat, Franz Josef Strauss Allee 11, D-93053 Regensburg, Germany
关键词
sncRNA; molecular beacon; piRNA; live cell; imaging; fluorescence; MESSENGER-RNA; LA; PHOSPHORYLATION; EXPRESSION; FRAGMENTS; TRANSPORT; ANTIGEN; LOOP;
D O I
10.1515/hsz-2023-0185
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Small non-coding RNAs (sncRNA) are involved in many steps of the gene expression cascade and regulate processing and expression of mRNAs by the formation of ribonucleoprotein complexes (RNP) such as the RNA-induced silencing complex (RISC). By analyzing small RNA Seq data sets, we identified a sncRNA annotated as piR-hsa-1254, which is likely derived from the 3'-end of 7SL RNA2 (RN7SL2), herein referred to as snc7SL RNA. The 7SL RNA is an abundant long non-coding RNA polymerase III transcript and serves as structural component of the cytoplasmic signal recognition particle (SRP). To evaluate a potential functional role of snc7SL RNA, we aimed to define its cellular localization by live cell imaging. Therefore, a Molecular Beacon (MB)-based method was established to compare the subcellular localization of snc7SL RNA with its precursor 7SL RNA. We designed and characterized several MBs in vitro and tested those by live cell fluorescence microscopy. Using a multiplex approach, we show that 7SL RNA localizes mainly to the endoplasmic reticulum (ER), as expected for the SRP, whereas snc7SL RNA predominately localizes to the nucleus. This finding suggests a fundamentally different function of 7SL RNA and its derivate snc7SL RNA.
引用
收藏
页码:1123 / 1136
页数:14
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