Research objectives and content
Over the last several years considerable progress has been made in understanding cell cycle controlling mechanisms. The identification of the negative growth regulator p53 and pl6ink4a and their involvement a; tumor suppressor proteins caused a major breakthrough in cell cycle research. In contrast to p53, very little is known about the extracellular or intracellular signals leading to the induction of p1 6-mediated grows! arrest. So far only oncogenic Ras has been shown to be a mediator of pl6ink4a-induced G1-growth arrest together with increased p53 in primary-like cells. Therefore, it is of outstanding interest to identify (novel' genes and gene products involved in oncogenic Ras-mediated pl6ink4a and p53 induction resulting insenescent-like growth arrest. The approach of retroviral transduction of a specially designed vector facilitates the selection of highly Ras-expressing cells by FACS-Analysis of green fluorescent protein coexpressing a normalized cDNA library.The identification of mediators involved in the Ras-induced G1-growth arrest will make a significanl contribution to the general understanding of both normal growth regulation and tumorigenesis.
Training content (objective, benefit and expected impact)
The objective of this training is to broaden the methodological repertoire of molecular ,,state of the art' techniques, in particular the use of the retroviral transduction method of primary cells. The handling of such retroviral constructs with an oncogenic Ras-fused green-fluorescent protein (GFP) as a selection marker for FACS-analysis can be easily adopted to other biologically relevant questions providing a powerful tool for future projects.
Links with industry / industrial relevance (22)