[tt] [Synthetic Biology] RNA visualization in live bacterial cells using fluorescent protein complementation.

[Eugen Leitl]

----- Forwarded message from Drew Endy <endy at MIT.EDU> -----

From: Drew Endy <endy at MIT.EDU>
Date: Wed, 23 May 2007 19:22:54 -0400
To: discuss at syntheticbiology.org
Subject: [Synthetic Biology] RNA visualization in live bacterial cells using
	fluorescent protein complementation.
X-Mailer: Apple Mail (2.752.3)

Another interesting article pointed out to me by Tom Knight.   Drew

Nat Methods. 2007 May;4(5):421-7. Epub 2007 Apr 1

RNA visualization in live bacterial cells using fluorescent protein  
complementation.

         * Valencia-Burton M,
         * McCullough RM,
         * Cantor CR,
         * Broude NE.

Center for Advanced Biotechnology and Department of Biomedical  
Engineering, Boston University, 36 Cummington St., Boston,  
Massachusetts 02215, USA.

     We describe a technique for the detection and localization of  
RNA transcripts in living cells. The method is based on fluorescent- 
protein complementation regulated by the interaction of a split RNA- 
binding protein with its corresponding RNA aptamer. In our design,  
the RNA-binding protein is the eukaryotic initiation factor 4A  
(eIF4A). eIF4A is dissected into two fragments, and each fragment is  
fused to split fragments of the enhanced green fluorescent protein  
(EGFP). Coexpression of the two protein fusions in the presence of a  
transcript containing eIF4A-interacting RNA aptamer resulted in the  
restoration of EGFP fluorescence in Escherichia coli cells. We also  
applied this technique to the visualization of an aptamer-tagged mRNA  
and 5S ribosomal RNA (rRNA). We observed distinct spatial and  
temporal changes in fluorescence within single cells, reflecting the  
nature of the transcript.

     PMID: 17401371 [PubMed - in process]

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