Down-regulation of gene expression in Gram-positive bacteria using RNA based techniques with emphasi

Project leader(s)   Kristin Hegstad Dahl, Researcher, Reference Centre for Detection of Antimicrobial Resistance (K-res)

Co-workers / Project staff
Arnfinn Sundsfjord, Professor Dept. of Microbiology and Virology University of Tromsø (UiTø) and K-res UNN Johanna E. Sollid, Associate professor Dept. of Microbiology and Virology UiTø Eirik W. Lundblad, Post doc. Dept. of Molecular Biotechnology, UiTø Anne-Merethe Hanssen, Post doc. Dept. of Microbiology and Virology UiTø Eva K. Bjørkeng, PhD student Dept. of Microbiology and Virology UiTø
Maria Sangvik, Research ass. K-res, UNN

Project outline
“Shoot the messenger”-strategies is a new innovative tool in functional genomic research of both bacteria and mammalian cells. This approach also holds promise for the development of a new class of therapeutics using RNA-molecules to silence unwanted gene expression in vivo. We will explore and use the capabilities of the endogenous, essential ribonuclease RNase P to target, cleave and inactivate specific mRNAs in our research efforts to understand the evolution, spread and persistence of pathogenic lineages of antimicrobial resistant Gram-positive bacteria. A successful technique may also have a therapeutic potential by targeting expression of antimicrobial resistance, virulence factors and essential elements for bacterial growth.

A functional genetic knockdown technique to down-regulate gene expression in Gram-positive bacteria using external guide sequences (EGS) will be developed through the identification of RNaseP substrate recognition and preferences. EGSs are anti-sense oligos that bind sequence-specifically to mRNA of selected target genes. This binding mimics natural RNase P targets in the cell and induces RNase P-mediated mRNA cleavage. Anti-sense technology offers an attractive alternative for microbial genomics exploring essential genes and clustered regions as it allows subtle control of gene expression rather than knockout.

Collaborators / Partners
National/international collaborator(s)
The molecular biology of RNA group with 1) Professor Steinar Johansens RNA lab (Dept. of Molecular Biotechnology, IMB), 2) Professor Trond Flægstads group (Dept. of Pediatrics, IKM), and 3) Professor Nils Peder Willassens Protein lab (Dept. of Molecular Biotechnology, IMB), UiT.
Dr. Roland K Hartmann, Philipps-Universität Marburg, Germany.
Dr. Rob JL Willems, University Medical Center Utrecht, The Netherlands.

Funding
Strategic Thematic Collaboration Project Funding at the Faculty of Medicine, UiT.
Helse-Nord RHF

Publications
None so far, the project was started July 2004.