Compositions comprising alternating 2′-modified nucleosides for use in gene modulation
The present invention provides compositions comprising at least one oligomeric compound comprising an alternating motif and further include a region that is complementary to a nucleic acid target. The compositions are useful for targeting selected nucleic acid molecules and modulating the expression of one or more genes. In preferred embodiments the compositions of the present invention hybridize to a portion of a target RNA resulting in loss of normal function of the target RNA. The present invention also provides methods for modulating gene expression.
1. A method of reducing target messenger RNA in a cell comprising contacting the cell with a composition comprising first and second chemically synthesized oligomeric compounds, wherein:
the first oligomeric compound is fully complementary to and capable of hybridizing to the second oligomeric compound and to the target messenger RNA;
each of the first and second oligomeric compounds independently comprises from 17 to 23 nucleosides; and
the first oligomeric compound comprises at least one motif selected from F(SF) n S nn where n is from 2 to about 20, nn is 0 or 1, one of F and S is a 2′-F modified nucleoside and the other of F and S is a 2′-O—CH 3 modified nucleoside;
thereby reducing the target messenger RNA in the cell.
2. The method of claim 1 wherein each F is a 2′-O—CH 3 modified nucleoside.
3. The method of claim 1 wherein each F is a 2′-F modified nucleoside.
4. The method of claim 1 wherein said first oligomeric compound further comprises a 5′-phosphate group.
5. The method of claim 1 wherein said second oligomeric compound further comprises a 5′-phosphate group.
6. The method of claim 1 wherein each of said first and said second oligomeric compounds independently, comprise a 5′-phosphate group.
7. The method of claim 1 wherein said first oligomeric compound comprises a 3′-terminal OH group.
8. The method of claim 1 wherein said first and said second oligomeric compounds comprise only phosphodiester internucleoside linking groups.
9. The method of claim 1 wherein said first and said second oligomeric compounds comprise only phosphorothioate internucleoside linking groups.
10. The method of claim 1 wherein said first oligomeric compound comprises only phosphodiester internucleoside linking groups and said second oligomeric compound comprises only phosphorothioate internucleoside linking groups; or said first oligomeric compound comprises only phosphorothioate internucleoside linking groups and said second oligomeric compound comprises only phosphodiester internucleoside linking groups.
11. The method of claim 1 wherein said first oligomeric compound comprises only phosphorothioate internucleoside linking groups and said second oligomeric compound comprises only phosphodiester internucleoside linking groups.
12. The method of claim 1 wherein said first and said second oligomeric compounds comprise internucleoside linking groups that are each independently selected from phosphodiester and phosphorothioate internucleoside linking groups.
13. The method of claim 1 wherein said first and said second oligomeric compounds comprise internucleoside linking groups that are each independently selected from phosphodiester, phosphorothioate, chiral phosphorothioate, phosphorodithioate, phosphotriester, aminoalkylphosphotriester, methyl phosphonate, alkyl phosphonate, 5′-alkylene phosphonate, chiral phosphonate, phosphinate, phosphoramidate, 3′-amino phosphoramidate, aminoalkylphosphoramidate, thionophosphoramidate, thionoalkylphosphonate, thionoalkylphosphotriester, selenophosphate and boranophosphate.
14. The method of claim 1 further comprising one conjugate group.
15. The method of claim 1 wherein at least one of said first and said second oligomeric compounds further comprises at least one conjugate group attached at the 3′-end or the 5′-end.
16. The method of claim 1 wherein at least one of said first and said second oligomeric compounds further comprises at least one terminal cap moiety attached at the 3′-end or the 5′-end or one terminal cap moiety attached at the 3′-end and a second terminal cap moiety attached at the 5′-end.
17. The method of claim 16 wherein each of said terminal cap moieties is an inverted deoxy abasic moiety.
18. The method of claim 16 wherein said second oligomeric compound is a sense strand comprising a terminal cap moiety at 3′-end, the 5′-end or one terminal cap at the 3′-end and a second terminal cap at the 5′-end.
19. The method of claim 18 wherein said terminal cap moiety is an inverted deoxy abasic moiety.
20. The method of claim 1 wherein said first and said second oligomeric compounds are a complementary pair of siRNA oligonucleotides.
21. The method of claim 1 wherein said first oligomeric compound is an antisense oligonucleotide.