Nucleic acid molecules inducing RNA interference, and uses thereof
The present invention relates to an RNAi-inducing nucleic acid molecule having a new structure and the use thereof, and more particularly to a novel nucleic acid molecule having a structure comprising a first strand, which is 24-121 nt in length and comprises a region complementary to a target nucleic acid, and a second strand which is 13-21 nt in length and has a region that binds complementarily to the region of the first strand, which is complementary to the target nucleic acid, so that the nucleic acid molecule inhibits the expression of a target gene with increased efficiency, and to a method of inhibiting the expression of a target gene using the nucleic acid molecule. The nucleic acid molecule structure of the present invention increases the efficiency with which the nucleic acid molecule inhibits the target gene. Alternatively, the nucleic acid molecule of the present invention can either increase the ability of the siRNA to bind to the target gene or cause synergistic cleavage, by introduction of antisense DNA, antisense RNA, ribozyme or DNAzyme, thereby increasing the efficiency with which the nucleic acid molecule inhibits the target gene. In addition, when the nucleic acid molecule according to the present invention is used, the efficiency with which the target gene is inhibited can be maintained for an extended period of time. Accordingly, the RNAi-inducing nucleic acid molecule of the present invention can be effectively used for the treatment of cancer or viral infection in place of conventional siRNA molecules.
1. A long antisense siRNA (lasiRNA) molecule comprising:
a first nucleic acid strand of 26-31 nucleotides (nt) in length and that is 100% complementary to a target nucleic acid; and
a second nucleic acid strand of 16 nt in length that binds complementarily to the first nucleic acid strand, such that the first nucleic acid strand has a double-stranded region of 16 nt in length to which the second nucleic acid strand binds and a single-stranded region of 10 nt to 15 nt in length to which the second nucleic acid strand does not bind, and wherein the 5′ end of the first nucleic acid strand and the 3′ end of the second nucleic acid strand form a blunt end.
2. The lasiRNA molecule of claim 1 , wherein the first nucleic acid strand is of 31 nt in length.
3. The lasiRNA molecule of claim 1 , wherein the target nucleic acid is a messenger RNA (mRNA).
4. The lasiRNA molecule of claim 1 , wherein the lasiRNA molecule comprises a chemical modification.
5. The lasiRNA molecule of claim 4 , wherein the chemical modification comprises a replacement of the hydroxyl group at position 2′ of ribose of at least one nucleotide included in the lasiRNA molecule by any one of a hydrogen atom, a fluorine atom, an —O-alkyl group, and an amino group.
6. The lasiRNA molecule of claim 4 , wherein the chemical modification comprises a replacement of the phosphate backbone of at least one nucleotide included in the lasiRNA molecule by any one of a phosphorothioate form, phosphorodithioate form, alkylphosphonate form, phosphoroamidate form, and boranophosphate form.
7. The lasiRNA molecule of claim 4 , wherein the chemical modification comprises a replacement of at least one nucleotide included in the lasiRNA molecule by any one of LNA (locked nucleic acid), UNS (unlocked nucleic acid) morpholino, and PNA (peptide nucleic acid).
8. The lasiRNA molecule of claim 4 , wherein the chemical modification comprises a lipid, cell penetrating peptide, or cell-targeting ligand bound to the lasiRNA molecule.
9. The lasiRNA molecule of claim 1 , wherein a cell delivery vehicle is bound to the lasiRNA molecule.
10. The lasiRNA molecule of claim 9 , wherein the cell delivery vehicle is selected from cationic polymers, lipids, cell penetrating peptides, and cell-targeting ligands.
11. The lasiRNA molecule of claim 1 , wherein the target gene is a tumor-related gene.
12. The lasiRNA molecule of claim 11 , wherein the tumor-related gene is any one of KRAS, Wnt-1, Hec1, Survivin, Livin, Bcl-2, XIAP, Mdm2, EGF, EGFR, VEGF, VEGFR, Mcl-1, IGF1R, Akt1, Grp78, STAT3, STAT5a, β-catenin, WISP1, and c-myc.
13. The lasiRNA molecule of claim 2 , wherein the lasiRNA molecule comprises a chemical modification.
14. The lasiRNA molecule of claim 13 , wherein the chemical modification comprises a replacement of the hydroxyl group at position 2′ of ribose of at least one nucleotide included in the lasiRNA molecule by any one of a hydrogen atom, a fluorine atom, an —O-alkyl group, and an amino group.
15. The lasiRNA molecule of claim 13 , wherein the chemical modification comprises a replacement of the phosphate backbone of at least one nucleotide included in the lasiRNA molecule by any one of a phosphorothioate form, phosphorodithioate form, alkylphosphonate form, phosphoroamidate form, and boranophosphate form.
16. The lasiRNA molecule of claim 13 , wherein the chemical modification comprises a replacement of at least one nucleotide included in the lasiRNA molecule by any one of LNA (locked nucleic acid), UNS (unlocked nucleic acid) morpholino, and PNA (peptide nucleic acid).
17. The lasiRNA molecule of claim 13 , wherein the chemical modification comprises a lipid, cell penetrating peptide, or cell targeting ligand bound to the lasiRNA molecule.
18. The lasiRNA molecule of claim 2 , wherein a cell delivery vehicle is bound to the lasiRNA molecule.
19. The lasiRNA molecule of claim 18 , wherein the cell delivery vehicle is selected from cationic polymers, lipids, cell penetrating peptides, and cell-targeting ligands.