Saarbrücken, Saarland, Germany
In my master‘s thesis, I am focussing on characterizing the interaction between The Nuclear Pore Complex and the spliceosome. To achieve this, I employ advanced techniques such as Fluorescence Microscopy and biochemical assays like Co-Immunoprecipation.
I used CellOracle to infer Gene Regulatory Networks in a PBMC dataset. CellOracle builds GRNs using scATAC-seq data to construct a base GRN and scRNA-seq data to refine strong network connections. CellOracle extracts information about key players in the network through graph-based network analysis.
Eukaryotes use splicing to remove non-coding introns from mRNA. This reaction is catalysed by the spliceosome and involves a lot of proteins. One protein, currently detected, is RNA helices Aquarius. Aquarius binds pre-mRNA within a pentameric intron-binding complex. Studies could show in vitro that one aquarius mutant (K829A) cause splicing deficiencies. My project should confirm this data in vivo. Therefor, I generate the mutants and transfect HEK293T cells.
This internship aimed at understanding the extracellular and intracellular factors of collective cell migration (also called flocking motion). In the past, the research group found out that over expressing GTPase Rab5A in breast epithelial cell lines (MCF10A, MCF10DCIS.com) cause reawaken of a quiescent/jammed cell sheet and unjamming. We stably transduced MCF10DCIS.com with a new plasmid, containing a Tet-on expression system. Moreover, we investigated the role of mechanical properties in collective cell migration. Stiffness describes the resistance of a material to stress. High stiffness means high resistance to any deformation. It was found out that cells preferably move towards stiffer material, a concept called durotaxis. We used an established protocol to induce flocking in HaCaT keratinocytes and seeded cells on PDMS well plates with known stiffness. Interestingly, cells move significantly faster on stiffer material. With HaCaT FUCCI cells, which enables tracking of cell cycle state, we monitored the proliferation on different stiffnesses and upon different conditions. Also cell proliferation seemed to be positively affected by stiffness.