Objectif The objective of this project is suppression of the insulin-like growth factor I (IGF-I) or its receptor (IGF-IR) biosynthesis by targeting their messenger RNA with synthetic nucleic acids. The principal emphasis will be done on the recent and most perspective strategy of small interfering RNA (siRNA) and deoxyribozymes (DNAzymes). In order to increase stability and penetration of the RNA oligomers into cells different modification of RNA backbones and eventually bases will be done and effect of these modifications on synthetic nucleic acid reagent efficacy will be studied.siRNA and deoxyribozyme action will be compared to that of other inhibitors such as antisense oligonucleotides, antisense RNA (targeting mRNA) or triple helix-forming oligonucleotides (targeting directly genomic double-stranded DNA). The most effective inhibitor will be selected in these comparative researches.Then the interaction of these inhibitors with living cells in culture will be studied. We propose to attach to these agents different chemically or biologically active or fluorescent molecules for studies of their penetration and intracellular behavior. Synthetic peptides will be used to facilitate penetration of the oligonucleotides into cells, as well as to address the inhibitor either to cytoplasm, or to nucleus and other subcellular structures.In order to achieve the objectives of the project the following studies will be undertaken:- Chemical synthesis of small double-stranded RNA and deoxyribozymes with natural backbone and with substitutions by modified nucleotides (2'-O-methyl or LNA) and shortened ribozymes with modified oligonucleotide effectors. - Synthesis of antisense and triple helix-forming oligonucleotides as well as their fluorescent conjugates and molecular beacons. - Design and synthesis of peptides facilitating penetration of nucleic acids inside the cells and improving their intracellular distribution.- Synthesis of conjugates between modified nucleic acid-based reagents and penetration peptides or molecules stabilizing complex formation with the targets.- Studies of penetration of these conjugates into the living cells by fluorescent or confocal microscopy.- Studies of effects of these synthetic inhibitors on the IGF-I and IGF-IR gene expression in cell cultures.Expected output of the project is to obtain new potent sequence specific synthetic molecules capable to knockdown the expression of IGF-I or its receptor in order to inhibit the oncogenic transformation processes and potentially to cure the development of certain tumors. The results of the research may have a great impact not only into fundamental studies of cancerogenesis, but also into potential anti-cancer therapeutical applications based on artificial regulation of gene expression. Programme(s) IC-INTAS - International Association for the promotion of cooperation with scientists from the independent states of the former Soviet Union (INTAS), 1993- Thème(s) OPEN - OPEN Call Appel à propositions Data not available Régime de financement Data not available Coordinateur Muséum National d'Histoire Naturelle Contribution de l’UE Aucune donnée Adresse rue Cuvier 43 75231 Paris Cedex 05 France Voir sur la carte Liens Site web Opens in new window Coût total Aucune donnée Participants (4) Trier par ordre alphabétique Trier par contribution de l’UE Tout développer Tout réduire CNRS - Université Pierre et Marie Curie Paris 6 France Contribution de l’UE Aucune donnée Adresse place Jussieu 4 75252 Paris Cedex 05 Voir sur la carte Coût total Aucune donnée M. V. Lomonosov Moscow State University Russie Contribution de l’UE Aucune donnée Adresse Leninskie gory 119992 Moscow Voir sur la carte Coût total Aucune donnée Siberian Branch of Russian Academy of Sciences Novosibirsk Institute of Bioorganic Chemistry Russie Contribution de l’UE Aucune donnée Adresse Lavrentyev prospekt 8 630090 Novosibirsk Voir sur la carte Coût total Aucune donnée University of Southern Denmark Danemark Contribution de l’UE Aucune donnée Adresse Campusvej 55 5230 Odense M Voir sur la carte Coût total Aucune donnée