Schoenberger, Johannes and Hammes, Ulrich Z. and Dresselhaus, Thomas (2012) In vivo visualization of RNA in plants cells using the lambda N-22 system and a GATEWAY-compatible vector series for candidate RNAs. PLANT JOURNAL, 71 (1). pp. 173-181. ISSN 0960-7412,
Full text not available from this repository. (Request a copy)Abstract
The past decade has seen a tremendous increase in RNA research, which has demonstrated that RNAs are involved in many more processes than were previously thought. The dynamics of RNA synthesis towards their regulated activity requires the interplay of RNAs with numerous RNA binding proteins (RBPs). The localization of RNA, a mechanism for controlling translation in a spatial and temporal fashion, requires processing and assembly of RNA into transport granules in the nucleus, transport towards cytoplasmic destinations and regulation of its activity. Compared with animal model systems little is known about RNA dynamics and motility in plants. Commonly used methods to study RNA transport and localization are time-consuming, and require expensive equipment and a high level of experimental skill. Here, we introduce the ?N22 RNA stem-loop binding system for the in vivo visualization of RNA in plant cells. The ?N22 system consists of two components: the ?N22 RNA binding peptide and the corresponding box-B stem loops. We generated fusions of ?N22 to different fluorophores and a GATEWAY vector series for the simple fusion of any target RNA 5' or 3' to box-B stem loops. We show that the ?N22 system can be used to detect RNAs in transient expression assays, and that it offers advantages compared with the previously described MS2 system. Furthermore, the ?N22 system can be used in combination with the MS2 system to visualize different RNAs simultaneously in the same cell. The toolbox of vectors generated for both systems is easy to use and promises significant progress in our understanding of RNA transport and localization in plant cells.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | BICOID MESSENGER-RNA; BINDING PROTEIN RRM4; ENDOPLASMIC-RETICULUM; FLUORESCENT PROTEIN; LOCALIZATION; TRANSPORT; SEQUENCE; IDENTIFICATION; RECOGNITION; EXPRESSION; fluorescence microscopy; GATEWAY technology; RNA binding protein; RNA transport; technical advance; lambda N22 |
| Subjects: | 500 Science > 580 Botanical sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Pflanzenwissenschaften > Lehrstuhl für Zellbiologie und Pflanzenphysiologie (Prof. Dr. Klaus Grasser) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 11 May 2020 09:10 |
| Last Modified: | 11 May 2020 09:10 |
| URI: | https://pred.uni-regensburg.de/id/eprint/18542 |
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