Spatio-termporal analysis of transcription dynamics in vivo
Final Report Abstract
Altogether, the project was not as successful as we hoped when we applied for funding. There were two events that caused severe deviations from the intended plans - one positive and one negative. (i) We discovered relatively early a new and so far unknown mechanism of regulation for ongoing transcription processes. Since it appeared to us very promising, we invested a lot of time and resources into this topic. We succeeded to prove this mechanism, however, could not analyze it at a molecular scale and could not identify any protein responsible for the observed effects. (ii) We were finally not successful in establishing super resolution STORM microscopy using the salivary gland cells. This was also due to the problem of the complex molecular gland surroundings. Finally, however, we succeeded to acquire super resolution image data using an Airyscan detector from the BR transcription sites using expansion microscopy. However, this work is not yet mature enough for publication.
Publications
- 2014. Transcription regulation during stable elongation by a reversible halt of RNA polymerase II. Mol Biol Cell. 25: 2190-8
Kaminski, T.P., J.P. Siebrasse and U. Kubitscheck
(See online at https://doi.org/10.1091/mbc.E14-02-0755) - 2015. Labelling and imaging of single endogenous messenger RNA particles in vivo. J. Cell Sci 128(20):3695-706
Spille, J.-H. and U. Kubitscheck
- 2017. Kinetics of transport through the nuclear pore complex. Semin. Cell Dev. Biol. 68: 18-26
Kubitscheck U., and Siebrasse, J.P.
(See online at https://doi.org/10.1016/j.semcdb.2017.06.016)
