Cel The objective of the present project is to develop new double-stranded DNA-binding ligands based on different conjugates between triple helix-forming oligonucleotides (TFO), minor groove binders (MGB), intercalators and inhibitors of topoisomerases I and II for specific regulation of gene expression. Their physico-chemical and biological properties will be studied, including their affinity to double-stranded DNA and stabilization of both canonical and unusual DNA structures such as triple helices. The sequence specific damage of DNA will be achieved by recruiting cellular enzymes topoisomerases by topoisomerase inhibitors conjugated to TFO and/or MGB acting as sequence-specific nucleases.In order to achieve the objectives of the project the following studies will be undertaken:- Synthesis of sequence-specific minor-groove binding ligands using pyrrole, imidazole, furan, thiazole benzimidazole, benzoxazole, indole and trimethoxyindole rings as building blocks;- Synthesis of covalent homo- and heterodimers of oligomeric MGB in head-to-tail, tail-to-tail and head-to-head orientations either by preliminary synthesis with the aim to conjugate them to oligonucleotides or by direct attachment of two MGB molecules to the same terminal phosphate of oligonucleotides. Studies of interaction of MGB and their conjugates with DNA;- Molecular modelling of the complexes and conjugates;- Synthesis of conjugates between minor groove binding ligands, triplex-forming oligonucleotides with natural and modified ribosophosphate backbone (locked nucleic acids - LNA, 2'-O-methyl oligoribonucleotides) and/or inhibitors of topoisomerases I and II such as camptothecin, rebeccamycin or etoposide. Thermal melting, foot printing and electrophoretic studies of duplex and triplex formation with target nucleic acids, stabilization of these structures and studies of affinity of synthetic ligands to double-stranded DNA;- Analysis of specific cleavage of DNA by TFO and/or MGB conjugates with topoisomerase inhibitors in the presence of topoisomerase I and II. Determination of the preferred sites of ligand binding on DNA using the DNase I footprinting technique, alkylation and the photo-induced and topoisomerase-induced cleavage of target molecules;- Testing of the synthetic ligands and their various conjugates as potential artificial sequence-specific nucleases and inhibitors of gene expression in vitro and in cell cultures.Expected output of the project is to obtain new potent sequence- and structure-specific DNA-binding molecules capable to recognize and stabilize double-stranded DNA structures, as well as artificial nucleases that may cleave DNA in desired sites by recruiting cellular enzymes and thus interfere with key cellular processes such as replication and transcription. These molecules could be proposed for potential design of new types of diagnostic and pharmacological agents. Program(-y) IC-INTAS - International Association for the promotion of cooperation with scientists from the independent states of the former Soviet Union (INTAS), 1993- Temat(-y) 4 - Life Sciences OPEN - OPEN Call Zaproszenie do składania wniosków Data not available System finansowania Data not available Koordynator Museum National d'Histoire Naturelle Wkład UE Brak danych Adres 43 rue Cuvier 75231 Paris cedex 05 Francja Zobacz na mapie Koszt całkowity Brak danych Uczestnicy (6) Sortuj alfabetycznie Sortuj według wkładu UE Rozwiń wszystko Zwiń wszystko Department of Chemistry Dania Wkład UE Brak danych Adres Campusvej 55 5230 Odense M Zobacz na mapie Koszt całkowity Brak danych Institute of Molecular Biology Rosja Wkład UE Brak danych Adres Novosibirsk Region 630559 Koltsovo Zobacz na mapie Koszt całkowity Brak danych Institute of Pharmaceutical Biotechnology Kazachstan Wkład UE Brak danych Adres 474456 Stepnogorsk Zobacz na mapie Koszt całkowity Brak danych Russian Academy of Sciences Rosja Wkład UE Brak danych Adres 8, Lavrentiev avenue 630090 Novosibirsk Zobacz na mapie Koszt całkowity Brak danych Russian Academy of Sciences Rosja Wkład UE Brak danych Adres Vavilov street 32 117984 Moscow Zobacz na mapie Koszt całkowity Brak danych UNIVERSITA DEGLI STUDI DI TRIESTE. Włochy Wkład UE Brak danych Adres Piazzale Europa 1 TRIESTE Zobacz na mapie Linki Strona internetowa Opens in new window Koszt całkowity Brak danych
