Background:
The main aim of our research is to understand the molecular and functional changes in astrocytic stem cells to study their regenerative potential. We are investigating the function of intermediate filament proteins, such as GFAP and Nestin, which are highly regulated cytoskeletal structures with a major role in astrocyte function. To elucidate the role of the filaments in the stem cell capacity, we aim to modulate their expression in neurogenic astrocyte cell lines. Gene silencing using RNA interference with short hairpins is a powerful tool to modify these cells. This approach allows us to investigate the role of GFAP and Nestin in stemness.
Aim:
This student project focuses on knockdown of Nestin in stem cell lines. This includes production of the expression vectors, validation of the knockdown efficiency and analysis of the effect of loss of Nestin on stem cell characteristics.
Methods:
Molecular Biology methods: Cloning (including restriction digest and analysis, ligation and transformation in E. coli), DNA and RNA amplification. Cell biology: Cell culture and cell transfections.
Contact:
Dr. Elly Hol (e.hol@nin.knaw.nl)
Drs. Regina Kanski (r.kanski@nin.knaw.nl)