METHOD FOR CIRCULARIZING OLIGONUCLEOTIDES AROUND A DOUBLE STRANDED NUCLEIC ACID, RESULTING STRUCTURES AND USES THEREOF

摘要

The invention concerns a method for circularizing an oligonucleotide which consists in: incubating a double stranded nucleic acid comprising a target sequence, with a single stranded oligonucleotide comprising a central part capable of being fixed on said target sequence, bound by spacer segments to 5' and 3' terminal sequences, said sequences being: either a) complementary to the sequence of an oligonucleotide matrix; or x) partly complementary to each other, so as to be capable of being hybridized to each other forming a double strand while preserving a single stranded end, which is capable of being hybridized with the terminal sequence of another oligonucleotide, called loop-oligonucleotide, which has a hairpin structure, with one single stranded part and a folded part forming a double strand; contacting, in the first case a) the complex formed with said oligonucleotide matrix, in conditions enabling the hybridization of the 3' and 5' ends, adjacently, with the matrix, and in the case x), contacting the complex formed with said loop-oligonucleotide, in conditions enabling, through hybridization of the free ends of the single stranded oligonucleotides and of the loop-oligonucleotide, the juxtaposition of the 3' and 5' ends of said oligonucleotides; ligating the hybridized ends so as to circularize the oligonucleotide around the double stranded nucleic acid and, optionally, isolating the circularized structure. The invention also concerns the applications and the resulting structures obtained for diagnostic and therapeutic purposes.