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RESEARCH - A. Rentmeister - Biomolecular label chemistry

 

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©Michael Kuhlmann
 

Research Profile
 
Our group primarily develops novel methods to selectively label biomolecules in living cells. Our main focus is labeling of mRNAs, which can localize to distinct subcellular regions in various cell types ranging from bacteria to neurons. To study the dynamics of mRNA localization and transport universal probes that allow to image mRNA in living cells are highly required. We harness and combine techniques from chemistry, molecular biology, and protein biochemistry to achieve selective labeling in cells. Our approaches aim at monitoring endogenous mRNA to increase the chances to study the transport and trafficking behavior of the target mRNA and not of artificial constructs.
 
1 - Chemo-enzymatic modification of mRNA
In a first strategy, we develop a new chemo-enzymatic method for mRNA modification in vitro. To achieve this, we engineer RNA-modifying enzymes, namely human Tgs1 (hTgs1) and Giardia lamblia Tgs2 (GlaTgs2) to incorporate functional groups suitable for bioorthogonal click chemistry into mRNA. These enzymes catalyze AdoMet-dependent methyl transfer to position N2 of the cap of RNAs. Using this approach and engineered enzyme variants we can now produce biotinylated and fluorescently labeled capped RNA in vitro. Our next goals are to further improve variants for interesting AdoMet analogues until conversion AdoMet-analogue is preferred converted over the native cofactor and to establish chemistry that can be used inside the cell.
 
2 - Engineering RNA-binding proteins
Our second strategy aims at expanding the repertoire of the Pum-homology domain of Homo sapiens Pumilio (HsPum1-HD). Pumilio is an RNA binding protein which binds to a stretch of 8 nucleotides of single stranded RNA in a sequence specific manner. We engineered a tetramolecular system that allows sequence specific RNA detection and discrimination between bound and unbound probe, which is essential for in vivo applications. We think that our strategy will contribute to improve current RNA imaging methods in living cells where it is imperative to discriminate between bound and unbound sample. Our system (i) lights up upon binding, (ii) shows minimal background, and (iii) could be entirely produced by the cellular machinery -essential properties for live cell imaging.
 

General publication list

Group members

If you are interested in a PhD position please contact
 
Prof. Dr. Andrea Rentmeister
Institut für Biochemie
WWU Münster
Wilhelm-Klemm-Strasse 2
D-48149 Münster
e-mail: a.rentmeister@uni-muenster.de

 


 

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