Our main research focus is the development and investigation of TAT (transactivator of transcription)- fusion proteins for neurodevelopmental and neurodegenerative disorders like RETT syndrome, Friedreich's Ataxia and Spinal Muscular Atrophy. The technology requires the synthesis of a fusion protein, linking the TAT transduction domain to the molecule of interest using a bacterial expression vector, followed by the purification of this fusion protein under either soluble or denaturing conditions. The purified fusion protein can be directly added to mammalian cell culture or injected in vivo into mice. Protein transduction occurs in a concentration-dependent manner, achieving maximum intracellular concentrations in less than 5 min, with nearly equal intracellular concentrations between all cells in the transduced population. Full-length TAT fusion proteins have been used to address a number of biological questions, relating to cell cycle progression, apoptosis, and cellular architecture [Dowdy SF et.al., Methods 2001].
Techniques, methods & infrastructure
Mouse models for RETT syndrome are used for in-vivo analysis and primary cells from human patients (e.g. fibroblasts) for in vitro analysis to investigate the properties of TAT fusion proteins. A broad spectrum of biochemical, cell biological, and imaging techniques are used (e.g. protein purification, immunohistochemistry, confocal microscopy, ECLIA, functional barriers,..)
- Steinkellner, H. et al., 2015. Detection of Survival Motor Neuron Protein in Buccal Cells Through Electrochemiluminescence-Based Assay. ASSAY and Drug Development Technologies, 13(3), pp.167-173. Available at: http://dx.doi.org/10.1089/adt.2015.635.
- Steinkellner, H. et al., 2014. Identification and molecular characterisation of a homozygous missense mutation in the ADAMTS10 gene in a patient with Weill-Marchesani syndrome. European Journal of Human Genetics, 23(9), pp.1186-1191. Available at: http://dx.doi.org/10.1038/ejhg.2014.264.
- Nachbauer, W. et al., 2011. Correlation of frataxin content in blood and skeletal muscle endorses frataxin as a biomarker in Friedreich ataxia. Movement Disorders, 26(10), pp.1935-1938. Available at: http://dx.doi.org/10.1002/mds.23789.
- Nachbauer, W. et al., 2011. Effects of Erythropoietin on Frataxin Levels and Mitochondrial Function in Friedreich Ataxia - a Dose-Response Trial. The Cerebellum, 10(4), pp.763-769. Available at: http://dx.doi.org/10.1007/s12311-011-0287-9.
- Steinkellner, H. et al., 2010. A high throughput electrochemiluminescence assay for the quantification of frataxin protein levels. Analytica Chimica Acta, 659(1-2), pp.129-132. Available at: http://dx.doi.org/10.1016/j.aca.2009.11.036.