Detail

Keywords

Blood-Brain Barrier; Cell-Penetrating Peptides; GABAergic Neurons; Rare Diseases; Rett Syndrome

Research group(s)

• LacconeLAB
  Research Area: Development and investigation of TAT (transactivator of transcription)- fusion proteins for neurodevelopmental and neurodegenerative disorders like RETT syndrome, Spinal Muscular Atrophy and Friedreich,
  Members:
  Hannes Steinkellner

Research interests

Our research focuses on developing and studying TAT (transactivator of transcription)-fusion proteins for neurodevelopmental and neurodegenerative disorders, including RETT syndrome, Friedreich's ataxia, and spinal muscular atrophy. This technology involves synthesizing a fusion protein by linking the TAT transduction domain to a target molecule using a bacterial expression vector, followed by purification under either soluble or denaturing conditions. The purified fusion protein can be directly applied to mammalian cell cultures or administered in vivo in mice.

Protein transduction occurs in a concentration-dependent manner, reaching maximum intracellular levels within five minutes and ensuring nearly uniform distribution across all transduced cells. Full-length TAT fusion proteins have been widely used to explore key biological processes such as 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,..)

Selected publications

1. Beribisky, A.V. et al. (2024) ‘MeCP2 is a naturally supercharged protein with cell membrane transduction capabilities’, Protein Science, 33(10). Available at: https://doi.org/10.1002/pro.5170.
2. Steinkellner, H. et al. (2022) ‘TAT-MeCP2 protein variants rescue disease phenotypes in human and mouse models of Rett syndrome’, International Journal of Biological Macromolecules, 209, pp. 972–983. Available at: https://doi.org/10.1016/j.ijbiomac.2022.04.080.
3. Steinkellner, H. et al. (2019) ‘An electrochemiluminescence based assay for quantitative detection of endogenous and exogenously applied MeCP2 protein variants’, Scientific Reports, 9(1). Available at: https://doi.org/10.1038/s41598-019-44372-3.
4. 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.
5. 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.