title: Rapid routes of synthesis of chemically reactive and highly radioactively labeled alpha- and beta-oligonucleotide derivatives for in vivo studies.
abstract: Development of the antisense oligonucleotide strategy for the regulation of gene expression in vivo poses several problems: the stability of oligonucleotides toward intracellular nucleases, labeling of oligonucleotides with high specific radioactivity, improvements of penetration of oligonucleotides into living cells, and enhancement of antisense action by coupling of chemically active groups. In the present paper synthesis of highly radioactively labeled [32P]- and [35S]oligonucleotide derivatives is described starting from both natural (beta) and nuclease-resistant (alpha) anomers of oligonucleotides. Conditions for preparative phosphorylation and thiophosphorylation suitable for oligonucleotides of various lengths, base composition, and anomeric forms were established. The stability of the phosphoramide bond under in vivo experimental conditions was checked. The methods of terminal phosphate chemical activation and terminal thiophosphate alkylation were applied to synthesize oligonucleotides equipped with hydrophobic, intercalating, alkylating, and photoactivatable groups. In the case of porphyrin-oligonucleotide conjugates, a series of new monofunctional porphyrin derivatives bearing a free aliphatic amino group was developed.
authors.affiliation: Laboratoire de Biophysique, Muséum National d'Histoire Naturelle, Paris, France.
authors.forename: A S
authors.initials: AS
authors.lastname: Boutorine
authors.forename: T
authors.initials: T
authors.lastname: Le Doan
authors.forename: J P
authors.initials: JP
authors.lastname: Battioni
authors.forename: D
authors.initials: D
authors.lastname: Mansuy
authors.forename: D
authors.initials: D
authors.lastname: Dupré
authors.forename: C
authors.initials: C
authors.lastname: Hélène
mesh_terms: D000166:Acridines / Q000138:chemical synthesis; D000255:Adenosine Triphosphate / Q000378:metabolism; D000477:Alkylating Agents / Q000138:chemical synthesis; D001483:Base Sequence; D055598:Chemical Phenomena; D002621:Chemistry; D002784:Cholesterol / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D005092:Exonucleases / Q000378:metabolism; D007553:Isotope Labeling; D007700:Kinetics; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D011166:Porphyrins / Q000138:chemical synthesis; D013462:Sulfur Radioisotopes
chemical_list: D000166:Acridines; D000477:Alkylating Agents; D004220:Disulfides; D009841:Oligonucleotides; D010761:Phosphorus Radioisotopes; D011166:Porphyrins; D013462:Sulfur Radioisotopes; D000255:Adenosine Triphosphate; D002784:Cholesterol; D005092:Exonucleases