Quantitative Analysis of Gene Regulation

• Unité de recherche: INNOSUISSE
• Numéro de projet: 7867.1;4 LSPP-LS
• Titre du projet anglais: Quantitative Analysis of Gene Regulation

Description succincte (Allemand)

Quantitative Analysis of Gene Regulation

Description succincte (Anglais)

Quantitative Analysis of Gene Regulation

Résumé des résultats (Abstract) (Allemand)

The project aims at the identification of gene regulatory elements along the chromosome which are target sites for binding transcription factors (TF). The background for this approach is the discovery of master control genes (Gehring) which in a hierarchical fashion control the expression of a large set of subordinate target genes and specify a given programme of organogenesis in insects as well as mammals. The development of fluorescence correlation spectroscopy and single molecule detection (Rigler) is already being used for gene expression analysis (Gnothis) and offers a unique method to identify specific protein-DNA complexes which are a prerequisit for gene regulation. The approach will be based on fluorescence tagging of TFs by Green Fluorescent Protein (GFP) and chromosomal DNA fragments with a small molecular weight dye using special fluorescently labelled primers. The analysis will be carried out in liquid microwell arrays developed by Gnothis which will be optimized for protein-DNA analysis. Large segments of chromosomes will be screened for TF binding sites using overlapping DNA segments of about 1-4 kb length obtained by PCR amplification. For this purpose only the TF has to be fluorescently tagged.The quantitative analysis of the in vitro target sites will be complemented subsequently by in vivo studies using chromatin co-immunoprecipitation. It will add a new dimension to our understanding of gene regulation at the transcriptional level.