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US5473060A Expired / Lapsed US ⏱️ 0.0 years remaining

Oligonucleotide clamps having diagnostic applications

Assignee: Lynx Therapeutics Inc
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Filing: 1993-07-02
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Granted: 1995-12-05
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Expires: 2013-07-02
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Abstract

Abstract Compounds referred to herein as oligonucleotide clamps are provided that stably bind to target polynucleotides in a sequence-specific manner. The oligonucleotide clamps comprise one or more oligonucleotide moieties capable of specifically binding to a target polynucleotide and one or more pairs of binding moieties covalently linked to the oligonucleotide moieties. In accordance with the invention, upon annealing of the oligonucleotide moieties to the target polynucleotide, the binding moieties of a pair are brought into juxtaposition so that they form a stable covalent or non-covalent linkage or complex. The interaction of the binding moieties of the one or more pairs effectively clamps the specifically annealed oligonucleotide moieties to the target polynucleotide.

Inventors

  • 👤 Sergei M. Gryaznov
  • 👤 David H. Lloyd

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-09 19:42
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/113; C07; C07H; C07H21/00; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12N2310/00; C12N2310/10; C12N2310/15; C12N2310/152; C12N2310/30; C12N2310/31; C12N2310/315; C12N2310/318; C12N2310/33; C12N2310/334; C12N2310/3341; C12N2310/35; C12N2310/351; C12N2310/3515

Claims Analysis

Total Claims: 15
Claim 1
Independent Claim 270 words
An oligonucleotide clamp for specifically hybridizing to a target polynucleotide, the oligonucleotide clamp comprising: (a) at least one oligonucleotide moiety capable of specifically hybridizing to the target polynucleotide with a Watson-Crick binding component and a Hoogsteen- or a reverse Hoogsteen-binding component; or at least two oligonucleotide moieties designated as OL1 and OL2 linked to a hinge region designated as G wherein at least one oligonucleotide moiety has a Watson-Crick binding component and at least one oligonucleotide moiety has a Hoogsteen- or a reverse Hoogsteen-binding component; and (b) at least one pair of non-oligonucleotide binding moieties, each pair of said binding moieties comprising a first binding moiety and a second binding moiety, the first binding moiety being covalently linked to an oligonucleotide moiety and the second binding moiety being covalently linked to an oligonucleotide moiety, wherein a stable covalent or non-covalent linkage is formed between the first binding moiety and the second binding moiety of the pair when the first and second binding moieties of the pair are brought into juxtaposition by the specific hybridization to the target polynucleotide of at least one or at least two oligonucleotide moieties, wherein said clamp has the formula: X--OL1--G--OL2--Y wherein: OL1 and OL2 are oligonucleotides specific for said target polynucleotide; G is a hinge region which links OL1 to OL2 so as to permit specific hybridization of OL1 and OL2 to their respective target polynucleotides; and X and Y are non-oligonucleotide binding moieties such that X and Y form a stable covalent or non-covalent linkage or complex whenever they are brought into juxtaposition by the hybridization of OL1 and OL2 to said target polynucleotide.
Claim 2
Dependent Claim 270 words
An oligonucleotide clamp for specifically hybridizing to a target polynucleotide, the oligonucleotide clamp comprising: (a) at least one oligonucleotide moiety capable of specifically hybridizing to the target polynucleotide with a Watson-Crick binding component and a Hoogsteen- or a reverse Hoogsteen-binding component; or at least two oligonucleotide moieties designated as OL1 and OL2 linked to a hinge region designated as G wherein at least one oligonucleotide moiety has a Watson-Crick binding component and at least one oligonucleotide moiety has a Hoogsteen- or a reverse Hoogsteen-binding component; and (b) at least one pair of non-oligonucleotide binding moieties, each pair of said binding moieties comprising a first binding moiety and a second binding moiety, the first binding moiety being covalently linked to an oligonucleotide moiety and the second binding moiety being covalently linked to an oligonucleotide moiety, wherein a stable covalent or non-covalent linkage is formed between the first binding moiety and the second binding moiety of the pair when the first and second binding moieties of the pair are brought into juxtaposition by the specific hybridization to the target polynucleotide of at least one or at least two oligonucleotide moieties, wherein said clamp has the formula: X--OL1--G--OL2--Y wherein: OL1 and OL2 are oligonucleotides specific for said target polynucleotide; G is a hinge region which links OL1 to OL2 so as to permit specific hybridization of OL1 and OL2 to their respective target polynucleotides; and X and Y are non-oligonucleotide binding moieties such that X and Y form a stable covalent or non-covalent linkage or complex whenever they are brought into juxtaposition by the hybridization of OL1 and OL2 to said target polynucleotide.
Claim 3
Dependent Claim 23 words
The oligonucleotide clamp of claim 1 further comprising one or more hinge regions for covalently linking two or more of said oligonucleotide moieties.
Claim 4
Dependent Claim 79 words
The oligonucleotide clamp of claim 1 wherein: OL1 and OL2 have the same length and are from 6 to about 40 nucleotides in length: and G is defined by the formula: (M--L).sub.n wherein: n is between 1 and 10; L is a linking group; and M is a straight chain, cyclic, or branched organic molecular structure containing from 1 to 20 carbon atoms and from 0 to 10 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur.
Claim 5
Dependent Claim 59 words
The oligonucleotide clamp of claim 3 wherein: M is an alkyl, alkoxy, alkenyl, or aryl containing from 1 to 16 carbon atoms; it heterocyclic having from 3 to 8 carbon atoms and from 1 to 3 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur; a glycosyl; or a nucleosidyl; and L is a phosphorus(V) linking group.
Claim 6
Dependent Claim 52 words
The oligonucleotide clamp of claim 4 wherein: M is an alkyl, alkoxy, alkenyl, or aryl containing from 1 to 8 carbon atoms; a glycosyl; or a nucleosidyl; L is selected from the group consisting of phosphodiester, phosphotdester, methyl phosphonate, ethyl phosphonate, phosphorothioate, phosphorodithioate, and phosphoramidate; and n is from 4 to 8.
Claim 7
Dependent Claim 14 words
The oligonucleotide clamp of claim 5 wherein X and Y form a hydrophobic complex.
Claim 8
Dependent Claim 27 words
The oligonucleotide clamp of claim 6 wherein X and Y are selected from the group consisting of alkanes, fatty acids, fatty alcohols, steroids, waxes, and fat-soluble vitamins.
Claim 9
Dependent Claim 28 words
The oligonucleotide clamp of claim 7 wherein X and Y are each a perhydrocyclopentanophenanthrene having from 19 to 30 carbon atoms and from 0 to 6 oxygen atoms.
Claim 10
Dependent Claim 13 words
The oligonucleotide clamp of claim 8 wherein X and Y are each cholesterol.
Claim 11
Dependent Claim 14 words
The oligonucleotide clamp of claim 5 wherein X and Y form a covalent linkage.
Claim 12
Dependent Claim 31 words
The oligonucleotide clamp of claim 10 wherein X is selected from the group consisting of phosphorothioate and phosphorodithioate and wherein Y is selected from the group consisting of haloacyl- or haloalkylamino.
Claim 13
Dependent Claim 12 words
The oligonucleotide clamp of claim 10 wherein X and Y are thiophosphoryl.
Claim 14
Dependent Claim 14 words
The oligonucleotide clamp of claim 1, wherein said G is 4 to 8 nucleotides.
Claim 15
Dependent Claim 44 words
The oligonucleotide clamp of claim 1, wherein said OL1 is a Watson-Crick binding component and said OL2 is a Hoogsteen- or a reverse-Hoogsteen binding component; or said OL1 is a Hoogsteen- or a reverse-Hoogsteen binding component and said OL2 is a Watson-Crick binding component.

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1998-12-04 1999-06-05 1999-12-05 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 2002-12-04 2003-06-05 2003-12-05 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 2006-12-04 2007-06-05 2007-12-05 $7700.00 Window Closed / Lapsed

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