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Tiny Molecular Beacons for in vivo mRNA Detection.
MedLine Citation:
PMID:  21431739     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
The molecular beacon technology is an established approach for visualizing native mRNAs in living cells. These probes need to efficiently hybridize to accessible RNA regions in order to spatially and temporally resolve the dynamic steps of the RNA life cycle. A refined method using two computer algorithms, mfold and RNAstructure, is described for choosing shorter, more abundant target regions for molecular beacon binding. The probes are redesigned as small hairpins and are synthesized from 2'-O-methyl RNA/LNA chimeric nucleic acids. These tiny molecular beacons are stable in the cellular environment and have a high affinity for binding to target RNAs. The user-friendly synthesis protocol and ability to couple to a variety of fluorophores make tiny molecular beacons the optimal technology to detect less abundant, highly structured RNAs, as well as small RNAs, such as microRNAs. As an example, tiny chimeric molecular beacons were designed to target regions of oskar mRNA, microinjected into living Drosophila melanogaster oocytes and imaged via spinning disc confocal microscopy.
Authors:
Diana P Bratu; Irina E Catrina; Salvatore A E Marras
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  714     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2011  
Date Detail:
Created Date:  2011-03-24     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  141-57     Citation Subset:  IM    
Affiliation:
Biological Sciences Department, Hunter College, City University of New York, New York, NY, USA, bratu@genectr.hunter.cuny.edu.
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