Oligonucleotide duplexes comprising DNA-like and RNA-like nucleotides and uses thereof
View Patent ↗Novel oligonucleotide pairs which can form a duplex comprising one or more DNA-like nucleotides (e.g., 2′-sub-stituted arabinonucleotides (ANA)); in combination with one or more RNA-like nucleotides (e.g., 2′-substituted ribonucleotides (RNA) and/or locked nucleic acid nucleotides (LNA)), are disclosed. The use of such oligonucleotide duplexes, such as for silencing the expression of a nucleic acid or gene of interest using small interfering RNA (siRNA) technologies, is also disclosed.
1. A hybridizable oligonucleotide pair for forming a duplex comprising a sense strand having a 5′ end and an antisense strand complementary to the sense strand,
the sense strand comprising a region 19 to 23 residues in length complementary to the antisense strand and consisting of: a plurality of 2′F-ANA and 2′F-RNA nucleotides, wherein at least a portion of the 2′F-ANA and 2′F-RNA nucleotides are present in a first altimer and a second, smaller altimer where the first altimer is located closer to the 5′ end of the sense strand than the second altimer; and
the antisense strand comprising a region 19 to 23 residues in length complementary to the sense strand and consisting of: a plurality of 2′F-RNA nucleotides or RNA nucleotides.
2. The oligonucleotide pair of claim 1 , wherein the sense strand, the antisense strand, Of both, comprises an overhang at the 3′ end, the overhang comprising DNA, 2′F-ANA, 2′F-RNA.
3. The oligonucleotide pair of claim 1 , wherein at least one of the sense strand and the antisense strand is phosphorylated at the 5′ end.
4. A double-stranded siRNA-like molecule comprising the oligonucleotide pair of claim 1 .
5. An shRNA-like molecule comprising a double-stranded stem corresponding to the double-stranded siRNA-like molecule of claim 4 .
6. A composition comprising the oligonucleotide pair of claim 1 and a pharmaceutically acceptable carrier.
7. A method of degrading or decreasing the level of a target nucleic acid, or of decreasing the level of a polypeptide encoded by said target nucleic acid, in a cell, the method comprising contacting the cell with the oligonucleotide pair of claim wherein the sense strand of the oligonucleotide pair comprises a nucleobase sequence substantially identical to a nucleobase sequence of the target nucleic acid.
8. The oligonucleotide pair of claim 1 , wherein the region of the antisense strand consists of RNA nucleotides.
9. The oligonucleotide pair of claim 1 , wherein the first altimer is a 3:3 altimer and the second altimer is a 1:1 altimer.
10. The oligonucleotide pair of claim 1 , wherein the region of the antisense strand consists of 2′F-RNA nucleotides.
11. The oligonucleotide pair of claim 1 , wherein the region of the antisense strand consists of RNA nucleotides and 2′F-RNA nucleotides.
12. The oligonucleotide pair of claim 9 , wherein the 3:3 altimer is [(2′F-ANA) 3 (2′F-RNA) 3 ].
13. The oligonucleotide pair of claim 9 , wherein the 1:1 altimer is [(2′F-ANA)(2′F-RNA)].
14. The oligonucleotide pair of claim 1 , wherein the region of the sense strand consists of [(2F-ANA) 3 (2′F-RNA) 3 ] 2 [(2′F-ANA)(2′F-RNA)] 3 (2′F-ANA).
15. The oligonucleotide pair of claim 1 , having a modified sense strand which results in a higher molar ellipticity at 220 nm compared by corresponding unmodified sense and antisense pairs.
16. The oligonucleotide pair of claim 1 wherein:
the region of the sense strand consists of; [(2′F-ANA) 3 (2′F-RNA) 3 ] 2 [(2′F-ANA)(2′F-RNA)] 3 (2′F-ANA), and the region of the antisense strand consists of: (RNA) 19 ; and
(b) the region of the sense strand consists of: [(2′F-ANA) 3 (2′F-RNA) 3 ] 2 [(2′F-ANA)(2′F-RNA)] 3 (2′F-ANA), and the region of the antisense strand consists of: (2′F-RNA) 19 .