A simplified vector system for visualization of localized RNAs in Schizosaccharomyces pombe |
| |
| Authors: | Tomoko Takeuchi-Andoh Sayaka Ohba Yu Shinoda Ayako Fuchita Sachiko Hayashi Emi Nishiyoshi |
| |
| Institution: | 1. Junior College Division, Otsuma Women’s University, Tokyo, Japan;2. Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, Kumamoto, Japan |
| |
| Abstract: | RNA localization is an important event that is essential for the polarization and differentiation of a cell. Although several methods are currently used to detect localized RNAs, a simplified detection system has not yet been developed for Schizosaccharomyces pombe. In the present study, we describe a new vector system for the visualization of localized RNAs in S. pombe using a U1A-tag-GFP system. A pREP1-U1A-tag vector plasmid to express U1A-tagged RNA and a pREP2-U1AGFP plasmid to produce a U1A-GFP fusion protein were constructed for this system. Since the U1A-GFP protein binds U1A-tagged RNA, fluorescence is observed at the location of U1A-tagged RNA in cells expressing both of these. The nucleolar localization of U3 snoRNA was successfully detected using this system, and a novel RNA localized at the DNA region of the nucleus was found by screening localized RNAs. This system will accelerate the study of localized RNAs in S. pombe. |
| |
| Keywords: | S pombe RNA localization visualization nucleus |
|
|
