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 method for inhibiting expression of a target gene in a mammalian cell, the method comprising introducing an RNAi-inducing nucleic acid molecule into the cell, wherein the RNAi-inducing nucleic acid molecule comprises:
a first strand of 24-119 nt length comprising a region 100% complementary to a target nucleic acid, wherein the region 100% complementary to the target nucleic acid comprises the 19 most 5′ nucleic acids of the first strand; and
a second strand of 16 nt length that binds complementarily to the region of the first strand 100% complementary to the target nucleic acid,
wherein the second strand binds to the first strand such that the first strand has a double-stranded region to which the second strand binds and a single-stranded region to which the second strand does not bind, and wherein the 5′ end of the first strand and the 3′ end of the second strand form a blunt end.
2. The method of claim 1 , wherein the target nucleic acid is a messenger RNA (mRNA).
3. The method of claim 1 , wherein the RNAi-inducing nucleic acid molecule comprises a chemical modification.
4. The method of claim 3 , wherein the chemical modification comprises a replacement of the hydroxyl group at position 2′ of ribose of at least one nucleotide included in the nucleic acid molecule by any one of a hydrogen atom, a fluorine atom, an —O-alkyl group and an amino group.
5. The method of claim 3 , wherein the chemical modification comprises a replacement of the phosphate backbone of at least one nucleotide included in the nucleic acid molecule by any one of a phosphorothioate form, phosphorodithioate form, alkylphosphonate form, phosphoroamidate form and boranophosphate form.
6. The method of claim 3 , wherein the chemical modification comprises a replacement of at least one nucleotide included in the nucleic acid molecule by any one of LNA (locked nucleic acid), UNS (unlocked nucleic acid) morpholino and PNA (peptide nucleic acid).
7. The method of claim 3 , wherein the chemical modification comprises a lipid, cell penetrating peptide or cell targeting ligand bound to the RNAi-inducing nucleic acid molecule.
8. The method of claim 1 , wherein a cell delivery vehicle is bound to the RNAi inducing nucleic acid molecule.
9. The method of claim 8 , wherein the cell delivery vehicle is selected from cationic polymers, lipids, cell penetrating peptides and cell targeting ligands.
10. A method for inhibiting expression of a target gene in a mammalian cell, the method comprising expressing an RNAi-inducing nucleic acid molecule in the cell, wherein the RNAi-inducing nucleic acid molecule comprises:
a first strand of 24-119 nt length comprising a region 100% complementary to a target nucleic acid wherein the region 100% complementary to the target nucleic acid comprises the 19 most 5′ nucleic acids of the first strand; and
a second strand of 16 nt length that binds complementarily to the region of the first strand 100% complementary to the target nucleic acid,
wherein the second strand binds to the first strand such that the first strand has a double-stranded region to which the second strand binds and a single-stranded region to which the second strand does not bind, and wherein the 5′ end of the first strand and the 3′ end of the second strand form a blunt end.
11. The method of claim 1 , wherein the target gene is a tumor-related gene.
12. The method 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 method of claim 10 , wherein the target gene is a tumor-related gene.
14. The method of claim 13 , 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.
15. The method of claim 1 , wherein the first strand is of 26-31 nt length.
16. The method of claim 1 , wherein the first strand is of 31 nt length.