Double-stranded ribonucleic acid with increased effectiveness in an organism

1. A double-stranded ribonucleic acid (dsRNA), consisting of first and second single RNA strands, wherein the first single RNA strand is an antisense RNA strand, the second single RNA strand is a sense RNA strand, wherein the antisense RNA strand is complementary to a target gene or a portion thereof, the dsRNA having increased effectiveness in inhibiting the expression of a target gene by means of RNA interference, wherein the dsRNA comprises first and second double-stranded ends and at least one single-stranded overhang which is 2 to 4 nucleotides in length, wherein the unpaired nucleotide of the single-stranded overhang that is directly adjacent to the terminal nucleotide base pair comprises a purine base, wherein a single-stranded overhang is located at the 3′-end of the antisense strand, wherein the overhang comprises the sequence 5′-GC-3′; and wherein the terminal base pair of the first double-stranded end comprises a guanine-cytosine (G-C) base pair or the four consecutive terminal base pairs of the first double-stranded end comprises at least two G-C base pairs; wherein the terminal base pair of the second double-stranded end comprises a guanine-cytosine (G-C) base pair or the four consecutive terminal base pairs of the second double-stranded end comprises at least two G-C base pairs; wherein the region of the antisense strand that is complementary to the targer gene is 19-28 nucleotides in length excluding the following dsRNAs:

5′- CAGGACCUCGCCGCUGCAGACC-3′

(SEQ ID NO: 1)

3′-CGGUCCUGGAGCGGCGACGUCUGG-5′,

(SEQ ID NO: 2)

5′- UGCAGCUUCGAAGCCUCACAGA-3′

(SEQ ID NO: 27)

3′-CGACGUCGAAGCUUCGGAGUGU-5′,

(SEQ ID NO: 28)

and

5′- UGGGGAGAGAGUUCUGAGGAUU-3′

(SEQ ID NO: 29)

3′-CGACCCCUCUCUCAAGACUCCU-5′

(SEQ ID NO: 30).

2. The dsRNA of claim 1 , wherein each nucleotide overhang independently consists of 2 unpaired nucleotides.

3. The dsRNA of claim 1 , wherein at least half of the unpaired nucleotides comprise a purine base.

4. The dsRNA of claim 1 , wherein the unpaired nucleotide adjacent to the terminal nucleotide base pair comprises a guanine (G).

5. The dsRNA of claim 1 , wherein the unpaired nucleotide adjacent to the terminal nucleotide base pair comprises an adenine (A) base.

6. The dsRNA of claim 1 , wherein said nucleotide overhang comprising the sequence 5′-GC-3′ consists of the sequence 5′-GC-3′.

7. The dsRNA of claim 1 , wherein the region of the antisense strand that is complementary to the target gene is 19 to 24 nucleotides in length.

8. The dsRNA of claim 1 , wherein the antisense strand is 20 to 28 nucleotides in length.

9. The dsRNA of claim 1 , wherein the antisense strand is 21 nucleotides in length.

10. The dsRNA of claim 1 , wherein at least one of the RNA strands comprises at least one chemically modified nucleotide.

11. The dsRNA of claim 10 , wherein the chemically modified nucleotide comprises a non-natural base.

12. The dsRNA of claim 10 , wherein the chemically modified nucleotide comprises a 2′ modification.

13. The dsRNA of claim 12 , wherein the 2′modification is selected from the group consisting of a 2′-amino modification, a 2′-alkyl modification, a 2′-O -methyl modification, a 2′-O-ethyl modification, a 2′-O-propyl modification, a 2′-O-allyl modification, a 2′-O-aminoalkyl modification, and a 2′-deoxy-2′-fluoro modification.

14. A method for the targeted selection of a double-stranded ribonucleic acid (dsRNA), consisting of first and second single RNA strands, wherein the first single RNA strand is an antisense RNA strand, the second single RNA strand is a sense RNA strand, wherein the antisense RNA strand is complementary to a target gene or a portion thereof, the dsRNA having increased effectiveness in inhibiting the expression of a target gene by means of RNA interference, comprising the steps of:

(a) selecting a dsRNA comprising first and second double-stranded ends and at least one single-stranded overhang which is 2 to 4 nucleotides in length;

(b) selecting a dsRNA comprising first and second double-stranded ends, wherein the unpaired nucleotide of the single-stranded overhang that is directly adjacent to the terminal nucleotide base pair comprises a purine base, wherein a single-stranded overhang is located at the 3′-end of the antisense strand, and wherein the overhang comprises the sequence 5′-GC-3′; and

(c) selecting a dsRNA comprising first and second double-stranded ends, wherein terminal base pair of the first double-stranded end comprises a guanine-cytosine (G-C) base pair or the four consecutive terminal base pairs of the first double-stranded end comprises at least two G-C base pairs; wherein the terminal base pair of the second double-stranded end comprises a guanine-cytosine (G-C) base pair or the four consecutive terminal base pairs of the second double-stranded end comprises at least two G-C base pairs; wherein the region of the antisense strand that is complementary to the target gene is 19-28 nucleotides in length;

excluding the following dsRNAs:

5′- CAGGACCUCGCCGCUGCAGACC-3′

(SEQ ID NO: 1)

3′-CGGUCCUGGAGCGGCGACGUCUGG-5′,

(SEQ ID NO: 2)

5′- UGCAGCUUCGAAGCCUCACAGA-3′

(SEQ ID NO: 27)

3′-CGACGUCGAAGCUUCGGAGUGU-5′,

(SEQ ID NO: 28)

and

5′- UGGGGAGAGAGUUCUGAGGAUU-3′

(SEQ ID NO: 29)

3′-CGACCCCUCUCUCAAGACUCCU-5′

(SEQ ID NO: 30).

15. The method of claim 14 , wherein each nucleotide overhang independently consists of 2 unpaired nucleotides.

16. The methods of claim 14 , wherein at least half of the unpaired nucleotides comprise a purine base.

17. The method of claim 14 , wherein the unpaired nucleotide adjacent to the terminal nucleotide base pair comprises a guanine (G) base.

18. The method of claim 14 , wherein the unpaired nucleotide adjacent to the terminal nucleotide base pair comprises an adenine (A) base.

19. The method of claim 14 , wherein said nucleotide overhang comprising the sequence 5′-GC-3′ consists of the sequence 5′-GC-3′.

20. The method of claim 14 , wherein the region of the antisense strand that is complementary to the target gene is 19 to 24 nucleotides in length.

21. The method of claim 14 , wherein the antisense strand is 20 to 28 nucleotides in length.

22. The method of claim 14 , wherein the antisense strand is 21 nucleotides in length.

23. The method of claim 14 , wherein at least one of the RNA strands comprises at least one chemically modified nucleotide.

24. The method of claim 23 , wherein the chemically modified nucleotide comprises a non-natural base.

25. The methods of claim 23 , wherein the chemically modified nucleotide comprises a 2′ modification.

26. The method of claim 25 , wherein the 2′ modification is selected from the group consisting of a 2′-amino modification, a 2′-alkyl modification, and a 2′-O-methyl modification, a 2′-O-ethyl modification, a 2′-O-propyl modification, a 2′-O-allyl modification, a 2′-O-aminoalkyl modification, and a 2′-deoxy-2′-fluoro modification.

27. A pharmaceutical composition for inhibiting the expression of a target gene by means of RNA interference, comprising a dsRNA of claim 1 , or a salt, prodrug or hydrate thereof; and a pharmaceutically acceptable carrier.

28. A method for inhibiting the expression of a target gene in a cell, comprising:

(a) introducing into the cell a dsRNA of claim 1 , or a salt, prodrug or hydrate thereof; and

(b) maintaining the cell for a time sufficient to obtain degradation of a mRNA transcript of the target gene.

29. The method of claim 28 , wherein the cell is a mammalian cell.

30. The method of claim 29 , wherein the cell is a human cell.

31. The method of claim 28 , wherein the target gene is selected from the group consisting of 11-hyroxysteroid dehydrogenase-1, acetyl-CoA-carboxylase-2, acyl CoA:

DAG acyltransferase1, Adenosine A2 receptor, akt, AML-ETO, amyloid beta precursor protein (APP), ApoAl, ApoB, ApoM, APS (adaptor protein with pleckstrin homology and src homology 2 domains, a-synuclein, Aurora A, Aurora B, beta-1 integrin subunit, beta-amyloid converting enzyme (BACE), Bax, beta-catenin, Bcl2, Bcl-XL, Bcr- abl, caspase 8, caspase-3, C CR2, CD40, CD40L, cdk2, chk1, chk2, clottingfactorVII, collagen, CD132, CTLA4, cyclin E, Dhcr24, Dipeptidylpeptidase-IV, E-Cadherin, Eg5/KSP, EGF, EGFR1, EWS-Flil, FAS-fatty acid synthase, FoxA-3, FoxO-1, Fructose-1,6-bisphosphate, Glucose-6-phophate, GM3 synthase, HDAC (histone deacetylase 1-6,9), Her-2/erb2, HIF1, HMG CoA reductase, hormone sensitive lipase, huntingtin, IKK1, IKK2, LDLR, MDR1, Microsomal Triglyceride Transfer Protein, MMP1, MMP2, MMP9, MyD88, sodium voltage gated type X alpha polypeptide (NaVl.8), NFkB, p38 map kinase mitogen activated protein kinase, p85a regulatory subunit of PI3-kinase, PEPCK, plkl, PTEN, PTP-1B, PU.1, raf, ras, Resistin, SCAP, SERBP-2, SHIP-2, SMAD7, SREBP1C, STAT1, stearoyl-CoA desaturase-1, TERT, TGF-beta-1, TGF-beta-1R1, Topoisomerase I, Topoisomerase II, VEGF, VEGFR1, VEGFR2, VLA1, VLA4, and vanilloid receptor (VR1).