Patent ID: 7566535
Filing Date: 2009-07-28
Classification: A61P,C12N

Abstract:
1. A method of oligonucleotide-mediated targeted sequence alteration of a nucleic acid, the method comprising: combining a target nucleic acid in the presence of cellular repair proteins with a sequence-altering targeting oligonucleotide; and either adding lambda beta protein additionally to said combination or first contacting cells having said cellular repair proteins with an HDAC inhibitor or hydroxyurea; wherein said oligonucleotide-mediated targeted sequence alteration is dependent upon a cellular DNA mismatch repair mechanism; wherein said oligonucleotide is a single-stranded oligonucleotide 17-121 nucleotides in length, said oligonucleotide having an internally unduplexed domain of at least 8 contiguous deoxyribonucleotides, wherein said oligonucleotide is fully complementary in sequence to the sequence of a first strand of the nucleic acid target, but for one or more mismatches as between the sequences of said internally unduplexed deoxyribonucleotide domain and its complement on said target nucleic acid first strand, each of said mismatches positioned at least 8 nucleotides from said oligonucleotide's 5′ and 3′ terminal, and wherein said oligonucleotide has at least one terminal modification selected from the group consisting of: at least one terminal locked nucleic acid (LNA), at least one terminal 2′—O—Me base analog, at least one terminal phosphorothioate linkage, and at least three terminal phosphorothioate linkages.