The cytoskeleton plays a key role in cell migration. However, cytoskeleton dynamics are regulated by several molecules that are therefore fundamental for the process of cell migration. Intriguingly, recent reports as well as results from our group identified a novel role for Rab proteins, mainly known for their function as master regulators of intracellular transport, in the processes of cell migration (Borg et al., 2014; Margiotta et al., 2017). These findings open up a completely new and exciting scenario of investigation on how membrane trafficking and cytoskeleton are co-regulated, and which are the players implicated in regulation of tumor migration and invasion. Therefore, one of the goals of our laboratory is to identify and characterize how intracellular transport can regulate cytoskeleton reorganization and the correct execution of cell migration programs.
Project:
We have recently performed a screening to identify Rab proteins involved in cancer cell migration and their interaction partners. In this project, the master student will learn state-of-the art cellular and molecular biology techniques in order to elucidate the role of the proteins identified in the screening in cell migration, and the underlying molecular mechanisms. In more detail, several biochemical and imaging methods will be used to characterize the interactions between the identified proteins. Also, the master student will deplete different cancer cell lines for the proteins of interest and study the effects on cytoskeleton and cell motility using migration, invasion and chemotaxis assays together with live imaging techniques in order to unravel the role of these proteins in both intracellular transport and cell migration.
Methods:
This project will use techniques in biochemistry, cellular and molecular biology, including:
- recombinant DNA technology
- PCR
- protein expression and purification
- immunoprecipitation
- Western blotting
- cell culture
- cell transfection
- RNA interference
- migration, invasion and chemotaxis assays
- immunofluorescence and state-of-the-art microscopy techniques, including FRET (fluorescence resonance energy transfer) and PLA (Proximity ligation assay) to study protein-protein interaction, FRAP (fluorescence recovery after photobleaching), and use of different microscopes available at the NorMic imaging platform (http://www.mn.uio.no/ibv/english/research/about/infrastructure/imaging/)
- quantitative image analysis using softwares such as ImageJ, Imaris, etc.
Contact info:
Phone: 22854441
Room: 3626