Doktorand i Immunotechnology

This advert is to find a candidate to fill the position of early stage researcher ESR6 in the Marie Curie Initial Training Network (MC-ITN) called ReBirth. The reBIRTH consortium focuses on molecular mechanisms of age-associated pathologies including neurodegeneration and depression. The nine international groups and companies work together to identify stress-regulated molecules provoking neuronal death and hindering neurogenesis, and monitor the consequences of these processes in human brain. The concept is that intelligent targeting of stress signaling will act synergistically to improve brain function in depressive disorders and ageing. The consortium is described in more detail at: http://www.itn-rebirth.eu

The successful ESR6 candidate will work on the project aimed at characterizing and developing new ways to perturb NMDA receptor related signaling complexes, The aim is to identify novel druggable pathway(s) modulating neurogenesis and contributing to depression. This will be done primarily in the laboratories of Peter James in Lund (Sweden) and Michael Courtney in Kuopio (Finland) with additional stays other laboratories of the network. To address NMDA receptor signaling pathways and the required protein complexes, the candidate will learn RNAi screen approaches, mass spectrometry applications for investigation of protein complexes, cell-permeable competitor peptide design, high-content analysis and high-throughput microscopy.

Arbetsuppgifter
(i) The first aim is to identify novel components of protein complexes involved in NMDA receptor signaling. Two strategies for RNAi screening of NMDA receptor functions will be considered in order to identify candidate genes involved. i) A high-throughput approach using a large panel of selected synthetic siRNAs or RNAi plasmids. The former is efficiently transfected into the majority of neurons, while the latter requires co-transfection with fluorescence reporter constructs; ii) Transfection of a single RNAi pool, followed by selection. In parallel, key proteins already known to assemble NMDA receptor-related signaling complexes will be precipitated by antibody or peptide ligand and co-precipitating complexes will be analysed by mass-spectrometry. The RNAi results will be evaluated for candidate scaffolding molecules for subsequent rounds of coIP- mass spectrometry.
(ii) Once the constituents of protein complexes have been established, the manner in which they assemble is addressed so that peptides disrupting assembly can be generated and validated in the same NMDA signaling assays used for RNAi as well as more complex models. The proteins will be expressed and pairwise or higher-order interactions investigated using cell-free recombinants and cell-based assays. Interacting regions are determined by deletion mapping in cell-based assay, with final validation and affinity measurement in cell-free system. The most promising sequences are used to generate cell-permeable peptide inhibitors validated as above.

Ledigkungörelse:
http://admin.lu.se/o.o.i.s?id=22598&Dnr=571218&Type=S

English version:
http://admin.lu.se/o.o.i.s?id=22598&Dnr=571218&Type=E