DNA virus microRNA and methods for inhibiting same

Abstract

The invention relates to isolated nucleic acid molecules comprising the sequence of any one of the DNA virus microRNAs shown in Table A1. In another embodiment, the invention relates to single stranded DNA virus microRNA molecules and anti-DNA virus microRNA molecules.

1 - 66 . (canceled)  
     
     
         67 . An isolated single stranded DNA virus microRNA molecule comprising a minimum of ten moieties and a maximum of fifty moieties on a molecular backbone, the molecular backbone comprising backbone units, each moiety comprising a base bonded to a backbone unit wherein: 
 at least ten contiguous bases have the same sequence as any one of the sequence of bases in a DNA virus microRNA molecule shown in Table A1, except that up to thirty percent of the bases pairs may be wobble base pairs, and up to 10% of the contiguous bases are additions, deletions, mismatches, or combinations thereof; and    no more than fifty percent of the contiguous moieties contain deoxyribonuleotide backbone units.    
     
     
         68 . An isolated single stranded anti-DNA virus microRNA molecule comprising a minimum of ten moieties and a maximum of fifty moieties on a molecular backbone, the molecular backbone comprising backbone units, each moiety comprising a base bonded to a backbone unit, each base forming a Watson-Crick base pair with a complementary base wherein: 
 at least ten contiguous bases have a sequence complementary to a contiguous sequence of bases in the sequence of bases in any one of the DNA virus microRNA molecule shown in Table A1, except that up to thirty percent of the bases pairs may be wobble base pairs, and up to 10% of the contiguous bases are additions, deletions, mismatches, or combinations thereof;    no more than fifty percent of the contiguous moieties contain deoxyribonuleotide backbone units; and    the molecule is capable of inhibiting microRNP activity.    
     
     
         69 . A molecule according to  claim 67 , further comprising a sequence of bases at the 5′ end and/or sequence of bases at the 3′ end present in a hairpin precursor from which the DNA microRNA is derived or any fragment thereof.  
     
     
         70 . A molecule according to  claim 69 , wherein the hairpin precursor is any one of the hairpin precursor sequences shown in Table A1 or any fragment thereof.  
     
     
         71 . A molecule according to  claim 67 , wherein the molecule is modified for increased nuclease resistance.  
     
     
         72 . A molecule according to  claim 68 , wherein the moiety in the molecule at the position corresponding to position 11 of the microRNA is non-complementary.  
     
     
         73 . A molecule according to  claim 68 , wherein up to 5% of the contiguous moieties are non-complementary to the contiguous sequence of bases in the DNA virus microRNA.  
     
     
         74 . A molecule according to  claim 68  having any one of the anti-DNA virus microRNA sequences shown in Table G1.  
     
     
         75 . A molecule according to  claim 68 , wherein at least one of the moieties is a deoxyribonucleotide.  
     
     
         76 . A molecule according to  claim 75 , wherein the deoxyribonucleotide is a modified deoxyribonucleotide moiety.  
     
     
         77 . A molecule according to  claim 76 , wherein the modified deoxyribonucleotide is a phosphorothioate deoxyribonucleotide moiety.  
     
     
         78 . A molecule according to  claim 76 , wherein the modified deoxyribonucleotide is N′3, —N′5 phosphoroamidate deoxyribonucleotide moiety.  
     
     
         79 . A molecule according to  claim 68 , wherein at least one of the moieties is a ribonucleotide moiety.  
     
     
         80 . A molecule according to  claim 79 , wherein at least one of the moieties is a modified ribonucleotide moiety.  
     
     
         81 . A molecule according to  claim 79 , wherein the modified ribonucleotide is substituted at the 2′ position.  
     
     
         82 . A molecule according to  claim 81 , wherein the substituent at the 2′position is a C 1  to C 4  alkyl group.  
     
     
         83 . A molecule according to  claim 82 , wherein the alkyl group is methyl.  
     
     
         84 . A molecule according to  claim 82 , wherein the alkyl group is allyl.  
     
     
         85 . A molecule according to  claim 81 , wherein the substituent at the 2′position is a C 1  to C 4  alkoxy-C 1  to C 4  alkyl group.  
     
     
         86 . A molecule according to  claim 85 , wherein the C 1  to C 4  alkoxy-C 1  to C 4  alkyl group is methoxyethyl.  
     
     
         87 . A molecule according to  claim 80 , wherein the modified ribonucleotide has a methylene bridge between the 2′-oxygen atom and the 4′carbon atom.  
     
     
         88 . A molecule according to  claim 68 , wherein at least one of the moieties is a peptide nucleic acid moiety.  
     
     
         89 . A molecule according to  claim 68 , wherein at least one of the moieties is a 2′-fluororibonucleotide moiety.  
     
     
         90 . A molecule according to  claim 68 , wherein at least one of the moieties is a morpholino phosphoroamidate nucleotide moiety.  
     
     
         81 . A molecule according to  claim 68 , wherein at least one of the moieties is tricyclo nucleotide moiety.  
     
     
         92 . A molecule according to  claim 68 , wherein at least one of the moieties is a cyclohexene nucleotide moiety.  
     
     
         93 . A molecule according to  claim 68 , wherein the molecule comprises at least one modified moiety for increased nuclease resistance.  
     
     
         94 . A molecule according to  claim 68 , wherein the molecule comprises a nucleotide cap at the 5′ end, the 3′ end, or both.  
     
     
         95 . A molecule according to  claim 68 , wherein the molecule comprises a chemical cap at the 5′ end, the 3′ end, or both.  
     
     
         96 . An isolated nucleic acid molecule comprising the sequence of any one of the DNA virus microRNA shown in Table A1.  
     
     
         97 . A method for inhibiting microRNP activity in a cell, the microRNP comprising a DNA virus microRNA molecule, the DNA virus microRNA molecule comprising a sequences of bases complementary to the sequence of bases in a single stranded anti-DNA virus microRNA molecule, the method comprising introducing into the cell a single-stranded anti-DNA virus microRNA molecule comprising a sequence of a minimum of ten moieties and a maximum of fifty moieties on a molecular backbone, the molecular backbone comprising backbone units, each moiety comprising a base bonded to a backbone unit, each base forming a Watson-Crick base pair with a complementary base, wherein: 
 at least ten contiguous bases of the anti-DNA virus microRNA molecule are complementary to any one of the DNA virus microRNAs shown in Table A1, except that up to thirty percent of the bases may be substituted by wobble base pairs, and up to ten percent of the at least ten moieties are addition, deletions, mismatches, or combinations thereof; and    no more than fifty percent of the contiguous moieties contain deoxyribonuleotide backbone units.    
     
     
         98 . An isolated microRNP comprising an isolated nucleic acid molecule according to  claim 67.