2008 - 2014Supervisor : Professor Anne Willis - Director of the MRC Toxicology Unit
MRC Toxicology Unit
Leicester, UK
Investigation of Translational Regulation during Cold-Shock
In recent years a growing number of reports investigated the use of reduced temperature cultivation (cold-shock) to enhance production of recombinant protein, Previous studies have suggested that there are several mechanisms by which cold-shock induced changes in gene expression including a general reduction in transcription and translation, inhibition of RNA degradation, increasing transcription of specific target genes and increasing translation of specific mRNA .
In this context, the aim of the project is to enhance global and mRNA specific translation for improved recombinant protein expression in in vitro cultured mammalian cells.
The objectives of my project are : 1) Profiling of total cellular mRNA and polysomal mRNA during cold-shock; 2) Ribonomics analysis of mRNA interacting directly with the cold-shock protein CIRP; 3) Identification of regulatory regions in mRNA that enhance translation upon cold-shock and 4) Design of vectors and optimisation of cells conditions that enhance protein production during cold shock.
INSERM U858
- PhD student
2003 - 2008Research work carried out as part of the doctoral research in « Hormones, growth factors and vascular physiopathology » Laboratory (U858 INSERM, Pr AC. Prats) University of Toulouse (France).
• Title: Study of translational regulation of the Vascular Endothelial Growth Factor - A isoforms.
• Supervisors: Pr. H. Prats and Dr. C. Touriol
• Projects: Evaluation of the effect of regulatory elements such as internal ribosome entry site and upstream open reading frame on the VEGF-A isoforms translation initiation and their imbrications with alternative spliced sequences.
This project is designed towards understanding fundamental processes within the regulation mechanism of VEGF isoforms implicated in pathological condition.
TECHNICAL SKILLS
• Cellular biology : Cultures of cell lines: HeLa, COS, fibroblast and ES; bacterial cultures (E.Coli).
• Molecular biology: Construction of vector, cloning, electroporation, transfection (plasmids/ RNA), PCR, RNA extraction, RT-PCR, Northen-Blot, RNase protection assay, Southern Blot.
• Protein biochemistry: Preparation of cytosolic and nuclear fractions, immunoblot, immunoprecipitation, co-immunoprecipitation, UV cross linking.
UMR6218 CNRS
- Postgraduate Researcher
2002 - 2003• Research Project: Engineering a PREP-1 knockout inducible gene targeting vector in the context of Down’s syndrome
