Methods and compositions involving mirna and mirna inhibitor molecules

Abstract

The present invention concerns methods and compositions for introducing miRNA activity or function into cells using synthetic nucleic acid molecules. Moreover, the present invention concerns methods and compositions for identifying miRNAs with specific cellular functions that are relevant to therapeutic, diagnostic, and prognostic applications wherein synthetic miRNAs and/or miRNA inhibitors are used in library screening assays.

What is claimed is: 
     
         1 . A synthetic RNA molecule of between 17 and 125 residues in length comprising
 a) an miRNA region whose sequence from 5′ to 3′ is identical to a mature miRNA sequence, and   b) a complementary region whose sequence from 5′ to 3′ is between 60% and 100% complementary to the miRNA sequence.   
     
     
         2 . The RNA molecule of  claim 1 , further comprising one or more of the following:
 i) a replacement group for phosphate or hydroxyl of the nucleotide at the 5′ terminus of the RNA molecule;   ii) one or more sugar modifications in the first or last 1 to 6 residues of the complementary region; or,   iii) noncomplementarity between one or more nucleotides in the last 1 to 5 residues at the 3′ end of the complementary region and the corresponding nucleotides of the miRNA region.   
     
     
         3 . The RNA molecule of  claim 2 , comprising a replacement group for phosphate or hydroxyl of the nucleotide at the 5′ terminus of the RNA molecule. 
     
     
         4 . The RNA molecule of  claim 3 , wherein the replacement group is biotin, an amine group, a lower alkylamine group, an acetyl group, 2′oxygen-methyl (2′O-Me), 4,4′-dimethoxytrityl with oxygen (DMTO), fluoroscein, a thiol, or acridine. 
     
     
         5 . The RNA molecule of  claim 3 , wherein the phosphate of the nucleotide at the 5′ terminus of the RNA molecule has been replaced. 
     
     
         6 . The RNA molecule of  claim 3 , wherein the hydroxyl of the nucleotide at the 5′ terminus of the RNA molecule has been replaced. 
     
     
         7 . The RNA molecule of  claim 2 , comprising one or more sugar modifications in the first or last 1 to 6 residues of the complementary region. 
     
     
         8 . The RNA molecule of  claim 7 , wherein the sugar modification is a 2′O-Me modification. 
     
     
         9 . The RNA molecule of  claim 7 , comprising one or more sugar modifications in the first 1 to 6 residues of the complementary region. 
     
     
         10 . The RNA molecule of  claim 7 , comprising one or more sugar modifications in the last 1 to 6 residues of the complementary region. 
     
     
         11 . The RNA molecule of  claim 7 , comprising one or more sugar modifications in the first or last 2 to 4 residues of the complementary region. 
     
     
         12 . The RNA molecule of  claim 2 , comprising noncomplementarity between one or more nucleotides in the last 1 to 5 residues at the 3′ end of the complementary region and the corresponding nucleotides of the miRNA region. 
     
     
         13 . The RNA molecules of  claim 12 , wherein there is noncomplementarity with at least 2 nucleotides in the complementary region. 
     
     
         14 . The RNA molecule of  claim 2 , comprising at least two of i), ii), or iii). 
     
     
         15 . The RNA molecule of  claim 14 , comprising i), ii), and iii). 
     
     
         16 . The RNA molecule of  claim 1 , wherein the RNA molecule is a single polynucleotide. 
     
     
         17 . The RNA molecule of  claim 16 , further comprising a linker region between the miRNA region and the complementary region. 
     
     
         18 . The RNA molecule of  claim 17 , wherein the linker region is between 3 and 30 residues in length. 
     
     
         19 . The RNA molecule of  claim 1 , wherein the RNA molecule is double stranded. 
     
     
         20 . A library of 10 or more different RNA molecules of  claim 1 . 
     
     
         21 . A library of 10 or more different miRNA inhibitor molecules, wherein each miRNA inhibitor molecule is between about 17 to 25 nucleotides in length and comprises a 5′ to 3′ sequence that is at least 90% complementary to the 5′ to 3′ sequence of a mature miRNA. 
     
     
         22 . The library of  claim 21 , wherein the miRNA inhibitor molecules have a replacement group for the phosphate or hydroxyl of the nucleotide at the 5′ terminus of the molecule. 
     
     
         23 . The library of  claim 22 , wherein the replacement group is an amine group, a lower alkylamine group, an acetyl group, or 2′O-Me. 
     
     
         24 . The library of  claim 22 , wherein the phosphate of the nucleotide at the 5′ terminus of the miRNA inhibitor molecule has been replaced. 
     
     
         25 . The library of  claim 22 , wherein the hydroxyl of the nucleotide at the 5′ terminus of the miRNA inhibitor molecule has been replaced. 
     
     
         26 . The library of  claim 21 , wherein the miRNA inhibitor comprises DNA. 
     
     
         27 . A method of characterizing an miRNA activity or function in a cell comprising:
 a) introducing into one or more cells the RNA molecule of  claim 1 ;   b) comparing one or more characteristics of cell(s) having the RNA molecule with cells not having the RNA molecule.   
     
     
         28 . The method of  claim 27 , wherein at least five RNA molecules are introduced into different cells. 
     
     
         29 . The method of  claim 27 , further comprising assaying the cell for the presence of the RNA molecule. 
     
     
         30 . The method of  claim 27 , further comprising synthesizing the RNA molecule. 
     
     
         31 . The method of  claim 30 , wherein the RNA molecule further comprises one or more of the following:
 i) a replacement group for phosphate or hydroxyl of the nucleotide at the 5′ terminus of the RNA molecule;   ii) one or more sugar modifications in the first or last 1 to 6 residues of the complementary region; or,   iii) noncomplementarity between one or more nucleotides in the last 1 to 5 residues at the 3′ end of the complementary region and the corresponding nucleotides of the miRNA region.   
     
     
         32 . The method of  claim 31 , wherein the RNA molecule comprises a replacement group for phosphate or hydroxyl of the nucleotide at the 5′ terminus of the RNA molecule. 
     
     
         33 . The method of  claim 32 , wherein the replacement group is biotin, an amine group, a lower alkylamine group, an acetyl group, 2′O-Me, DMTO, fluoroscein, a thiol, or acridine. 
     
     
         34 . The method of  claim 32 , wherein the phosphate of the nucleotide at the 5′ terminus of the RNA molecule has been replaced. 
     
     
         35 . The method of  claim 32 , wherein the hydroxyl of the nucleotide at the 5′ terminus of the RNA molecule has been replaced. 
     
     
         36 . The method of  claim 31 , wherein the RNA molecule comprises one or more sugar modifications in the first or last 1 to 6 residues of the complementary region. 
     
     
         37 . The method of  claim 36 , wherein the sugar modification is a 2′O-Me modification. 
     
     
         38 . The method of  claim 36 , wherein the RNA molecule comprises one or more sugar modifications in the first 1 to 6 residues of the complementary region. 
     
     
         39 . The method of  claim 36 , wherein the RNA molecule comprises one or more sugar modifications in the last 1 to 6 residues of the complementary region. 
     
     
         40 . The method of  claim 36 , wherein the RNA molecule comprises one or more sugar modifications in the first or last 2 to 4 residues of the complementary region. 
     
     
         41 . The method of  claim 31 , wherein the RNA molecule comprises noncomplementarity between one or more nucleotides in the last 1 to 5 residues at the 3′ end of the complementary region and the corresponding nucleotides of the miRNA region. 
     
     
         42 . The method of  claim 41 , wherein there is noncomplementarity with at least 2 nucleotides in the complementary region. 
     
     
         43 . The method of  claim 31 , wherein the RNA molecule comprises at least two of i), ii), or iii). 
     
     
         44 . The method of  claim 43 , wherein the RNA molecule comprises i), ii), and iii). 
     
     
         45 . The method of  claim 27 , wherein the RNA molecule is a single polynucleotide. 
     
     
         46 . The method of  claim 45 , wherein the RNA molecule further comprises a linker region between the miRNA region and the complementary region. 
     
     
         47 . The method of  claim 46 , wherein the linker region is between 3 and 30 residues in length. 
     
     
         48 . The method of  claim 27 , wherein the RNA molecule is composed of two polynucleotides. 
     
     
         49 . The method of  claim 27 , wherein the RNA molecule is introduced into the cell by calcium phosphate transfection, lipid transfection, electroporation, microinjection, or injection. 
     
     
         50 . A method of reducing or eliminating activity of one or more miRNAs from a cell comprising:
 a) introducing into a cell an miRNA inhibitor that is between 17 and 25 nucleotides in length and comprises a 5′ to 3′ sequence that is at least 90% complementary to the 5′ to 3′ sequence of a mature miRNA.   
     
     
         51 . The method of  claim 50 , further comprising:
 b) comparing one or more characteristics of a cell having the miRNA inhibitor with a cell not having the miRNA inhibitor.   
     
     
         52 . The method of  claim 50 , further comprising assaying the cell for expression of the mature miRNA. 
     
     
         53 . The method of  claim 50 , further comprising synthesizing the miRNA inhibitor. 
     
     
         54 . The method of  claim 51 , wherein the miRNA inhibitor further comprises one or more of the following:
 i) a replacement group for phosphate or hydroxyl of the nucleotide at the 5′ terminus of the miRNA inhibitor; or   ii) one or more sugar modifications.   
     
     
         55 . The method of  claim 54 , wherein the miRNA inhibitor comprises a replacement group for phosphate or hydroxyl of the nucleotide at the 5′ terminus of the miRNA inhibitor. 
     
     
         56 . The method of  claim 55 , wherein the replacement group is biotin, an amine group, a lower alkylamine group, an acetyl group, 2′O-Me, DMTO, fluoroscein, a thiol, or acridine. 
     
     
         57 . The method of  claim 55 , wherein the phosphate of the nucleotide at the 5′ terminus of the miRNA inhibitor has been replaced. 
     
     
         58 . The method of  claim 55 , wherein the hydroxyl of the nucleotide at the 5′ terminus of the miRNA inhibitor has been replaced. 
     
     
         59 . The method of  claim 54 , wherein the miRNA inhibitor comprises one or more sugar modifications in the first or last 1 to 6 residues. 
     
     
         60 . The method of  claim 59 , wherein the sugar modification is a 2′O-Me modification. 
     
     
         61 . The method of  claim 59 , wherein the miRNA inhibitor comprises one or more sugar modifications in the first 1 to 6 residues of the complementary region. 
     
     
         62 . The method of  claim 59 , wherein the miRNA inhibitor comprises one or more sugar modifications in the last 1 to 6 residues of the complementary region. 
     
     
         63 . The method of  claim 59 , wherein the miRNA inhibitor comprises one or more sugar modifications in the first or last 2 to 4 residues of the complementary region. 
     
     
         64 . The method of  claim 54 , wherein the miRNA inhibitor comprises both i) and ii). 
     
     
         65 . The method of  claim 50 , wherein the miRNA inhibitor is a single polynucleotide. 
     
     
         66 . The method of  claim 50 , wherein the miRNA inhibitor is introduced into the cell by calcium phosphate transfection, lipid transfection, electroporation, microinjection, or injection. 
     
     
         67 . A method for reducing or inhibiting cell proliferation in a cell comprising introducing into the cell an effective amount of at least one
 i) miRNA inhibitor molecule that is between 17 and 25 nucleotides in length and comprises a 5′ to 3′ sequence that is at least 90% complementary to the 5′ to 3′ sequence of a mature miRNA selected from the group consisting of: mir-31, mir-92, mir-99a, mir-100, mir-125a, mir-129, mir-130a, mir-150, mir-187, miR-190, miR-191, miR-193, miR 204, mir-210, mir-211, mir-212, mir-213, mir-215, mir-216, mir-217, miR 218, mir-224, mir-292, mir-294, mir-320, mir-324, mir-325, mir-326, mir-330, mir-331, mir-338, mir-341, mir-369, and mir-370; or   ii) synthetic miRNA molecule of between 17 and 125 residues in length comprising an miRNA region whose sequence from 5′ to 3′ is identical to a mature miRNA sequence, and a complementary region whose sequence from 5′ to 3′ is between 60% and 100% complementary to the mature miRNA sequence, wherein the mature miRNA sequence is selected from the group consisting of: miR-15a, miR-16, miR 21, miR 24, miR-96, miR-101, miR-105, miR-124, miR-126, miR-142, miR-147, miR-192, miR-194, miR-206, miR-215, or miR-346.   
     
     
         68 . The method of  claim 67 , further comprising identifying a cell or a patient having the cell as in need of reducing or inhibiting cell proliferation. 
     
     
         69 . A method for inducing or increasing cell proliferation in a cell comprising introducing into the cell an effective amount of at least one
 i) miRNA inhibitor molecule that is between 17 and 25 nucleotides in length and comprises a 5′ to 3′ sequence that is at least 90% complementary to the 5′ to 3′ sequence of a mature miRNA selected from the group consisting of let7a-1, Let-7a, Let-7b, let7b-1, let7c, let7d, Let-7g, mir-9, mir-10a, mir-10b, mir-15a, mir-16, mir-21, mir-23a, mir-23b, mir-24, mir-25, mir-92, mir-95, mir-133a, mir-133a-2, mir-133b, mir-142, mir-152, mir-153, mir-155, mir-181a, mir-182, mir-183, mir-184, mir-186, mir-187, mir-191, mir-193, mir-194, mir-196, mir-199a-1, mir-200b, mir-204, mir-206, mir-211, mir-222, mir-223, mir-298, mir-328, mir-342, mir-371, and mir-412; or   ii) synthetic miRNA molecule of between 17 and 125 residues in length comprising an miRNA region whose sequence from 5′ to 3′ is identical to a mature miRNA sequence, and a complementary region whose sequence from 5′ to 3′ is between 60% and 100% complementary to the miRNA sequence selected from the group consisting of: Let-7a, Let-7b, Let-7c, Let-7d, Let-7g, miR-7, mir-9, miR-10a, miR-10b, miR-15a, miR-18, miR-19a, miR-17-3p, miR-20, miR-23b, mir-25, miR-26a, miR-26a, mir-30e-5p, mir-31, mir-32, mir-92, mir-93, miR-100, miR-125a, miR-125b, miR-126, mir-127, miR-128, miR-129, mir-130a, mir-135, mir-138, mir-139, miR-140, mir-141, mir-143, mir-145, mir-146, miR-150, mir-154, mir-155, mir-181a, miR-182, mir-186, miR-187, miR-188, mir-190, mir-191, mir-193, mir-194, mir-196, mir-197, mir-198, mir-199, mir-201, mir-204, mir-216, mir-218, miR-223, mir-293, miR-291-3p, miR-294, miR-295, miR-322, mir-333, mir-335, mir-338, mir-341, mir-350, mir-369, miR-373, mir-410, and mir-412.   
     
     
         70 . The method of  claim 69 , further comprising identifying a cell or a patient having the cell as in need of inducing or increasing cell proliferation. 
     
     
         71 . A method for reducing cell viability comprising introducing into one or more cells an effective amount of at least one
 i) miRNA inhibitor molecule that is between 17 and 25 nucleotides in length and comprises a 5′ to 3′ sequence that is at least 90% complementary to the 5′ to 3′ sequence of a mature miRNA selected from the group consisting of: miR-107, miR-133, miR-137, miR-152, miR-155, miR-181a, miR-191, miR-203, and miR-215; or   ii) synthetic miRNA molecule of between 17 and 125 residues in length comprising an miRNA region whose sequence from 5′ to 3′ is identical to a mature miRNA sequence, and a complementary region whose sequence from 5′ to 3′ is between 60% and 100% complementary to the miRNA sequence selected from the group consisting of: let-7a, let-7b, mir-1, mir-7, miR-10b, miR-17-3p, miR-19a, mir-23, mir-24, mir-27a, miR-29a, miR-30a-3p, mir-31, mir-32, miR-34a, miR-101, miR-107, miR-108, miR-122, mir-124, miR-133a, miR-134, miR-135, miR-139, mir-140, miR-141, miR-145, mir-150, mir-192, mir-193, mir-195, mir-206, mir-208, mir-210, mir-210, mir-292-3p, mir-293, mir-297, mir-299, mir-329, mir-337, mir-337, mir-345, mir-346, and mir-409.   
     
     
         72 . The method of  claim 71 , further comprising identifying a cell or a patient having the cell as in need of reducing cell viability. 
     
     
         73 . A method for increasing cell viability comprising introducing into one or more cells an effective amount of at least one
 i) miRNA inhibitor molecule that is between 17 and 25 nucleotides in length and comprises a 5′ to 3′ sequence that is at least 90% complementary to the 5′ to 3′ sequence of a mature miRNA selected from the group consisting of miR-7, miR-19a, miR-23, miR-24, miR-27a, miR-31, miR-32, miR-134, miR-140, miR-150, miR-192, and miR-193; or   ii) synthetic miRNA molecule of between 17 and 125 residues in length comprising an miRNA region whose sequence from 5′ to 3′ is identical to a mature miRNA sequence, and a complementary region whose sequence from 5′ to 3′ is between 60% and 100% complementary to the miRNA sequence selected from the group consisting of: let-7a, let-7b, let-7c, let-7d, let-7g, miR-10a, miR-10b, miR-15a, miR-17-3p, miR-18, miR-19a, miR-20, mir-23a, mir-23b, mir-24, miR-25, miR-26a, mir-32, miR-107, miR-125a, miR-126, mir-128, miR-129, miR-133, miR-137, mir-139, miR-143, miR-152, miR-155, miR-181a, miR-182, miR-191, miR-203, miR-215, and mir-331.   
     
     
         74 . The method of  claim 73 , further comprising identifying a cell or a patient having the cell as in need of increasing cell viability. 
     
     
         75 . A method for inducing apoptosis in a cell comprising introducing into the cell an effective amount of at least one
 i) miRNA inhibitor molecule that is between 17 and 25 nucleotides in length and comprises a 5′ to 3′ sequence that is at least 90% complementary to the 5′ to 3′ sequence of a mature miRNA selected from the group consisting of: miR-31 and miR-214; or   ii) synthetic miRNA molecule of between 17 and 125 residues in length comprising an miRNA region whose sequence from 5′ to 3′ is identical to a mature miRNA sequence, and a complementary region whose sequence from 5′ to 3′ is between 60% and 100% complementary to the miRNA sequence selected from the group consisting of: let-7b, let-7g, mir-1, mir-1d, mir-7, mir-10a, miR-10b, miR-17-3p, miR-19a, miR-28, miR-28, miR-28, miR-29a, miR-32, miR-34a, miR-122, mir-148, mir-149, mir-154, mir-184, mir-186, mir-188, mir-192, mir-195, mir-196, mir-199a, mir-204, mir-208, mir-210, mir-211, mir-212, mir-214, mir-215, mir-216, mir-217, mir-218, mir-293, mir-296, mir-299, mir-321, mir-328, and mir-344.   
     
     
         76 . The method of  claim 75 , further comprising identifying a cell or a patient having the cell as in need of inducing apoptosis. 
     
     
         77 . A method for inhibiting apoptosis in a cell comprising introducing into the cell an effective amount of at least one
 i) miRNA inhibitor molecule that is between 17 and 25 nucleotides in length and comprises a 5′ to 3′ sequence that is at least 90% complementary to the 5′ to 3′ sequence of a mature miRNA selected from the group consisting of: miR-7, miR-1d, miR-148, miR-195, miR-196, miR-199a, miR-204, miR-210, miR-211, miR-212, miR-215, miR-216, miR-218, miR-296, and miR-321; or   ii) synthetic miRNA molecule of between 17 and 125 residues in length comprising an miRNA region whose sequence from 5′ to 3′ is identical to a mature miRNA sequence, and a complementary region whose sequence from 5′ to 3′ is between 60% and 100% complementary to the miRNA sequence selected from the group consisting of: Let-7b, mir-21, mir-23b, mir-25, miR-26a, mir-28, mir-29a, mir-31, miR-32, mir-30a-3p, mir-34a, mir-96, miR-98, mir-100, mir-101, mir-105, mir-108, miR-125b, miR-126, mir-126, mir-128, mir-137, miR-143, miR-155, mir-207, mir-214, mir-216, mir-223, mir-292-3p, mir-328, mir-335, mir-340, mir-341, mir-367, mir-368, mir-380-3p, and mir-410.   
     
     
         78 . The method of  claim 77 , further comprising identifying a cell or a patient having the cell as in need of inhibiting apoptosis. 
     
     
         79 . A method for treating cancer in a subject comprising administering to the subject an effective amount of one or more synthetic miRNA molecules and/or miRNA inhibitors corresponding to miR-21, miR-126, miR-143, miR-145, miR-188, miR-200b, miR-219, or miR-331. 
     
     
         80 . The method of  claim 79 , wherein the cancer is breast cancer and the subject is administered 1) one or more miRNA inhibitors corresponding to miR-21, miR-15a, miR-16, miR-24, and/or miR-25, and/or 2) one or more miRNAs corresponding to miR-99, miR-100, miR-205, miR-197, miR-126, miR-143, miR-145 and/or miR-321. 
     
     
         81 . The method of  claim 79 , wherein the cancer is colon cancer and the subject is administered 1) one or more miRNA inhibitors corresponding to miR-21, miR-106, miR-189, miR-200b, miR-223, miR-224, miR-31, and/or miR-17; and/or 2) one or more miRNAs corresponding to miR-145, miR-143, miR-133, miR-342, miR-125a, miR-195, miR-30a, miR-10a, miR-130, miR-130a, miR-192, miR-194, miR-215, miR-144, miR-23, miR-26a, miR-126, miR-199a, miR-188, miR-331, and/or miR-21. 
     
     
         82 . The method of  claim 79 , wherein the cancer is thyroid cancer and the subject is administered 1) one or more miRNA inhibitors corresponding to miR-21 miR-125, miR-24, miR-200b, miR-29b, miR-221, miR-222, miR-224, miR-10a, and/or miR-183; and/or 2) one or more miRNAs corresponding to miR-145, miR-22, miR-331, miR-126, miR-30a, miR-199a, miR-223, and/or miR-321. 
     
     
         83 . The method of  claim 79 , wherein the cancer is lung cancer and the subject is administered 1) one or more miRNA inhibitors corresponding to miR-223, miR-106, miR-21, miR-200b, miR-321, miR-182, miR-183, miR-17, and/or miR-205; and/or 2) one or more miRNAs corresponding to miR-130a, miR-145, miR-126, miR-331, miR-342, miR-143, Let-7, miR-30a, miR-16, miR-26a, miR-125a, miR-29b, miR-24, miR-328, miR-201, miR-195, miR-22, miR-181a, miR-331, and/or miR-321. 
     
     
         84 . A method for treating cancer in a subject comprising administering to the subject an effective amount of one or more synthetic miRNA molecules and/or miRNA inhibitors corresponding to let-7, miR-10a, miR-16, miR-17, miR-21, miR-22, miR-23, miR-24, miR-26a, miR-29b, miR-30a, miR-106, miR-125a, miR-126, miR-130, miR130a, miR-133, miR-143, miR-144, miR-145, miR-181a, miR-182, miR-183, miR-188, miR-189, miR-219, miR-192, miR-194, miR-195, miR-199a, miR-200b, miR-201, miR-205, miR-215, miR-223, miR-224, miR-321, miR-328, miR-331, or miR-342. 
     
     
         85 . A method of treating cancer in a patient comprising administering to the patient the cancer therapeutic and an effective amount of at least one miRNA molecule that improves the efficacy of the cancer therapeutic or protects non-cancer cells. 
     
     
         86 . The method of  claim 85 , wherein the miRNA molecule enhances the efficacy of the cancer therapeutic and is selected from the group consisting of ambi-miR-7100, mir-28, mir-101, mir-124, mir-125a, mir-126, mir-132, mir-136, mir-147, mir-155, mir-182, mir-186, mir-202, mir-206, mir-216, mir-221, mir-224, mir-291, mir-292-3p, mir-297, mir-302, mir-337, mir-372, mir-373, and mir-376b. 
     
     
         87 . The method of  claim 85 , wherein the miRNA molecule protects non-cancer cells from the cancer therapeutic and is selected from the group consisting of mir-16, mir-24, mir-30a-3p, mir-125b, mir-152, mir-194, mir-197, mir-214, and mir-331. 
     
     
         88 . A method for treating lupus in a subject comprising administering to the subject an effective amount of 1) one or more miRNA inhibitors corresponding to miR-21, miR-223, and/or mir-342 expression; and/or 2) one or more miRNAs corresponding to miR-95, miR-105, miR-137, miR-186, miR-188, miR-199, miR-211, miR-215, mu-miR-290, miR-301, and/or miR-331. 
     
     
         89 . A method for treating a prion disease in a subject comprising administering to the subject an effective amount of 1) one or more miRNA inhibitors corresponding miR-7, miR-9-as, miR-16, miR-24, miR-26A, miR-27A, miR-130A, and/or miR-239 expression; and/or 2) one or more miRNAs corresponding to miR-95 and/or miR-135A. 
     
     
         90 . A method for preventing or treating ischemia in a subject comprising administering to the subject an effective amount of 1) one or more miRNA inhibitors corresponding to miR-28, miR-30A, miR-31, miR-138, miR-139, miR-140, miR-291-5P and/or miR-298 expression; and/or 2) one or more miRNAs corresponding to Let7F-2 and/or miR-16. 
     
     
         91 . A method for inducing transformation in a cell comprising administering to the cell an effective amount of at least one miRNA selected from the group consisting of mir-192, mir-198, and mir-199. 
     
     
         92 . A method for activating ERK in cells comprising providing or introducing into the cells an effective amount of one or more nucleic acids capable of being processed into a mature miRNA when it is inside the cell, wherein the mature miRNA is let-7, miR-19a, miR-25, miR-96, miR-125a, miR-134, miR-148, miR-152, miR-206, miR-207, miR-210, miR-212, miR-216, miR-217, miR-218, miR-223, mu-miR-294, mu-miR-295, miR-301, miR-328, mu-miR-329, miR-339, miR-370, or miR-372. 
     
     
         93 . A method for activating hTert in a cell comprising introducing into the cell an effective amount of one or more nucleic acids capable of being processed into a mature miRNA inside the cell, wherein the mature miRNA is miR-15a, miR-16, miR-21, mir-24, miR-26a, miR-92, miR-105, miR-125a, miR-125b, miR-128, mir-147, miR-195, miR-207, miR-224, mu-miR-295, mir-301, miR-337, mir-368, or mir-371. 
     
     
         94 . A method for identifying an miRNA that inhibits an hTert activating gene product comprising:
 a) introducing into a cell a candidate miRNA into a cell;   b) assaying the level of hTert expression or hTert activity in the cell, wherein a reduction in hTert expression or activity compared to a cell lacking the miRNA identifies the miRNA as a potential inhibitor of an hTert activating gene product.   
     
     
         95 . The method of  claim 94 , wherein the sequence of the candidate miRNA was previously evaluated for an ability to inhibit an hTert activating gene product. 
     
     
         96 . The method of calim 87.7, wherein the hTert activating gene product is selected from the group consisting of ACOX1, AKT1, APAF1, COX-5B, COX6, COX7B, CPOX, DUOX2, GPX1, GPX2, GPX4, LPO, MAPK1, MAPK4, MTCO1, NOX3, NOX5, PAOX, PPDX, PRKCA, PRKCD, and TNFRSF6. 
     
     
         97 . A method for inhibiting hTert in a cell comprising providing to the cell an effective amount of an miRNA selected from the group consisting of miR-26a, miR-147, mir-195, and mir-368. 
     
     
         98 . A method for inhibiting stimulation of Stat3 in a cell comprising providing to the cell an effective amount of an miRNA selected from the group consisting of mir-93, mir-100, mir-134, mir-99a, mir-103, mir-128, mir-129, mir-181b, mir-193, mir-197, mir-212, mir-218, mir-219, mir-302, mir-323, mir-324-3p, mir-325, mir-330, mir-331, mir-340, mmu-mir-350, mir-425, mir-491, mir-518f, and mir-520a*. 
     
     
         99 . A kit for evaluating miRNA molecules comprising, in suitable containers, at least five RNA molecules of  claim 1 . 
     
     
         100 . A kit for evaluating miRNA molecules comprising, in suitable containers, at least five miRNA inhibitor molecules, wherein each miRNA inhibitor molecule is between 17 and 25 nucleotides in length and comprises a 5′ to 3′ sequence that is at least 90% complementary to the 5′ to 3′ sequence of a mature miRNA. 
     
     
         101 . A kit comprising, in suitable containers:
 a) at least one synthetic miRNA molecule of between 17 and 125 residues in length comprising an miRNA region whose sequence from 5′ to 3′ is identical to a mature miRNA sequence, and a complementary region whose sequence from 5′ to 3′ is between 60% and 100% complementary to the miRNA sequence; and   b) at least one miRNA inhibitor between 17 and 25 nucleotides in length and comprising a 5′ to 3′ sequence that is at least 90% complementary to the 5′ to 3′ sequence of a mature miRNA.