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In vivo visualization of RNA in plants cells using the λN 22 system and a GATEWAY‐compatible vector series for candidate RNAs
Author(s) -
Schönberger Johannes,
Hammes Ulrich Z.,
Dresselhaus Thomas
Publication year - 2012
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2012.04923.x
Subject(s) - rna , computational biology , biology , riboswitch , microbiology and biotechnology , rna binding protein , translation (biology) , nucleic acid structure , non coding rna , genetics , messenger rna , gene
Summary 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 λN 22 RNA stem‐loop binding system for the in vivo visualization of RNA in plant cells. The λN 22 system consists of two components: the λN 22 RNA binding peptide and the corresponding box‐B stem loops. We generated fusions of λN 22 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 λN 22 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 λN 22 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.