US5176996A
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Method for making synthetic oligonucleotides which bind specifically to target sites on duplex DNA molecules, by forming a colinear triplex, the synthetic oligonucleotides and methods of use
Abstract A method for making synthetic oligonucleotides which bind to target sequences in a duplex DNA forming colinear triplexes by binding to the major groove. The method includes scanning genomic duplex DNA and identifying nucleotide target sequences of greater than about 20 nucleotides having either about at least 65% purine bases or about at least 65% pyrimidine bases; and synthesizing synthetic oligonucleotides complementary to identified target sequences. The synthetic oligonucleotides have a G when the complementary location in the DNA duplex has a GC base pair and have a T when the complementary location in the DNA duplex has an AT base pair. The synthetic oligonucleotides are oriented 5' to 4' and bind parallel or 3' to 5' and bind anti-parallel to the about at least 65% purine strand.
A method of making a synthetic oligonucleotide which binds to a target sequence in duplex DNA forming a collinear triplex by binding to the major groove, said method comprising the steps of: (a) identifying a duplex nucleotide target sequence of greater than about 20 nucleotides on each strand, said target sequence having either about at least 65% purine bases or about at least 65% pyrimidine bases on each strand; and (b) synthesizing said synthetic olignucleotide complementary to said identified target sequence, said synthetic oligonucleotide including a G and a T, wherein said synthetic oligonucleotide has a G when the complementary location in the DNA duplex has a GC base pair and has a T when the complementary location in the DNA duplex has an AT base pair.
Claim 2
Independent Claim
127 words
A method of making a synthetic oligonucleotide which binds to a target sequence in duplex DNA forming a collinear triplex by binding to the major groove, said method comprising the steps of: (a) identifying a duplex nucleotide target sequence of greater than about 20 nucleotides on each strand, said target sequence having either about at least 65% purine bases or about at least 65% pyrimidine bases on each strand; and (b) synthesizing said synthetic olignucleotide complementary to said identified target sequence, said synthetic oligonucleotide including a G and a T, wherein said synthetic oligonucleotide has a G when the complementary location in the DNA duplex has a GC base pair and has a T when the complementary location in the DNA duplex has an AT base pair.
Claim 3
Dependent Claim
39 words
The method of claim 1 wherein said synthetic oligonucleotide is oriented 3' to 5' and binds anti-parallel to the about at least 65% purine strand, wherein the orienting strand of the duplex DNA is the more purine rich strand.
Claim 4
Dependent Claim
39 words
The method of claim 1, wherein said synthetic oligonucleotide is oriented 5' to 3' and binds parallel to the about at least 65% purine strand, wherein the orienting strand of the duplex DNA is the more purine rich strand.