US5476928A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Modified nucleotides and polynucleotides and complexes form therefrom
Assignee: Yale University
•
Filing: 1992-02-26
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Granted: 1995-12-19
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Expires: 2012-02-26
Abstract
Abstract Compounds having the structure: ##STR1## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety covalently bonded to the C 1' -position of the sugar moiety, provided that when B is purine or 7-deazapurine, it is attached at the N 9 -position of the purine or 7-deazapurine and when B is pyrimidine, it is attached at the N 1 -position; wherein A represents a moiety consisting of at least three carbon atoms which is capable of forming a detectable complex with a polypeptide when the compound is incorporated into a double-stranded ribonucleic acid, deoxyribonucleic acid duplex, or DNA-RNA hybrid; wherein the dotted line represents a chemical linkage joining B and A, provided that if B is purine, the linkage is attached to the 8-position of the purine, if B is 7-deazapurine, the linkage is attached to the 7-position of the deazapurine, and if B is pyrimidine, the linkage is attached to the 5-position of the pyrimidine; and wherein each of x, y and z represents ##STR2## either directly, or when incorporated into oligo- and polynucleotides, provide probes which are widely useful. Applications include detection and localization of polynucleotide sequences in chromosomes, fixed cells, tissue sections, and cell extracts. Specific applications include chromosomal karyotyping, clinical diagnosis of nucleic acid-containing etiological agents, e.g. bacteria, viruses, or fungi, and diagnosis of genetic disorders.
Inventors
- 👤 David C. Ward
- 👤 Pennina R. Langer
- 👤 Alexander A. Waldrop, III
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 15:38
CPC Classifications
C; C07; C07H; C07H19/00; C07H19/02; C07H19/04; C07H19/16; C07H19/20; C07H21/00; C12; C12P; C12P19/00; C12P19/26; C12P19/28; C12P19/30; C12P19/34; C12Q; C12Q1/00; C12Q1/02; C12Q1/18; C12Q1/68; C12Q1/6804; C12Q1/6813; C12Q1/6816
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
207 words
A compound useful as a probe for detecting the presence or absence of a nucleic acid, said compound having the structure: ##STR26## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety suitable for incorporation into a polynucleotide and covalently bonded to the C 1 '-position of the sugar moiety, provided that when B is a purine or 7-deazapurine, the sugar moiety is attached at the N 9 -position of the purine or deazapurine, and when B is pyrimidine, the sugar moiety is attached at the N 1 position of the pyrimidine; wherein A represents at least three carbon atoms and an indicator molecule selected from the group consisting of fluorescent dyes, electron-dense reagents, enzymes which can be reacted with a substrate to produce a visually detectable reaction product, and radioisotopes; wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with detection of A; wherein if B is a purine, A is attached to the 8-position of the purine, if B is a 7-deazapurine, A is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A is attached to the 5-position of the pyrimidine; and wherein each of x, y and z represents: ##STR27##
Claim 2
Dependent Claim
207 words
A compound useful as a probe for detecting the presence or absence of a nucleic acid, said compound having the structure: ##STR26## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety suitable for incorporation into a polynucleotide and covalently bonded to the C 1 '-position of the sugar moiety, provided that when B is a purine or 7-deazapurine, the sugar moiety is attached at the N 9 -position of the purine or deazapurine, and when B is pyrimidine, the sugar moiety is attached at the N 1 position of the pyrimidine; wherein A represents at least three carbon atoms and an indicator molecule selected from the group consisting of fluorescent dyes, electron-dense reagents, enzymes which can be reacted with a substrate to produce a visually detectable reaction product, and radioisotopes; wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with detection of A; wherein if B is a purine, A is attached to the 8-position of the purine, if B is a 7-deazapurine, A is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A is attached to the 5-position of the pyrimidine; and wherein each of x, y and z represents: ##STR27##
Claim 3
Dependent Claim
234 words
A compound useful as a probe for detecting the presence or absence of a nucleic acid, said compound containing a nucleotide having the structure: ##STR28## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety covalently bonded to the C 1 '-position of the sugar moiety, provided that whenever B is purine or 7-deazapurine, the sugar moiety is attached at the N 9 -position of the purine or deazapurine, and whenever B is a pyrimidine, the sugar moiety is attached at the N 1 -position of the pyrimidine; wherein A represents at least three carbon atoms and an indicator molecule selected from the group consisting of fluorescent dyes, electron-dense reagents, enzymes which can be reacted with a substrate to produce a visually detectable reaction product and radioisotopes; wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with detection of A; wherein if B is a purine, A or the linkage group is attached to the 8-position of the purine, if B is a 7-deazapurine, A or the linkage group is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A or the linkage group is attached to the 5-position of the pyrimidine; wherein one of x and y represents ##STR29## and the other of x and y is absent or represents --OH or --H; and wherein z represents H-- or HO--.
Claim 4
Dependent Claim
234 words
A compound useful as a probe for detecting the presence or absence of a nucleic acid, said compound containing a nucleotide having the structure: ##STR28## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety covalently bonded to the C 1 '-position of the sugar moiety, provided that whenever B is purine or 7-deazapurine, the sugar moiety is attached at the N 9 -position of the purine or deazapurine, and whenever B is a pyrimidine, the sugar moiety is attached at the N 1 -position of the pyrimidine; wherein A represents at least three carbon atoms and an indicator molecule selected from the group consisting of fluorescent dyes, electron-dense reagents, enzymes which can be reacted with a substrate to produce a visually detectable reaction product and radioisotopes; wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with detection of A; wherein if B is a purine, A or the linkage group is attached to the 8-position of the purine, if B is a 7-deazapurine, A or the linkage group is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A or the linkage group is attached to the 5-position of the pyrimidine; wherein one of x and y represents ##STR29## and the other of x and y is absent or represents --OH or --H; and wherein z represents H-- or HO--.
Claim 5
Dependent Claim
223 words
A complex useful as a probe for detecting the presence or absence of a nucleic acid, said complex comprising a detectable polypeptide complexed with a compound having the structure: ##STR30## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety suitable for incorporation into a polynucleotide and covalently bonded to the C 1 '-position of the sugar moiety, provided that when B is a purine or 7-deazapurine, the sugar moiety is attached at the N 9 -position of the purine or deazapurine, and when B is pyrimidine, the sugar moiety is attached at the N 1 position of the pyrimidine; wherein A represents at least three carbon atoms, is capable of specifically complexing with the detectable polypeptide when A is linked to B, and represents a component of a signalling moiety capable of producing a detectable signal; wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with the characteristic ability of A to form said complex with the detectable polypeptide; wherein if B is a purine, A is attached to the 8-position of the purine, if B is a 7-deazapurine, A is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A is attached to the 5-position of the pyrimidine; and wherein each of x, y and z represents: ##STR31##
Claim 6
Dependent Claim
223 words
A complex useful as a probe for detecting the presence or absence of a nucleic acid, said complex comprising a detectable polypeptide complexed with a compound having the structure: ##STR30## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety suitable for incorporation into a polynucleotide and covalently bonded to the C 1 '-position of the sugar moiety, provided that when B is a purine or 7-deazapurine, the sugar moiety is attached at the N 9 -position of the purine or deazapurine, and when B is pyrimidine, the sugar moiety is attached at the N 1 position of the pyrimidine; wherein A represents at least three carbon atoms, is capable of specifically complexing with the detectable polypeptide when A is linked to B, and represents a component of a signalling moiety capable of producing a detectable signal; wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with the characteristic ability of A to form said complex with the detectable polypeptide; wherein if B is a purine, A is attached to the 8-position of the purine, if B is a 7-deazapurine, A is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A is attached to the 5-position of the pyrimidine; and wherein each of x, y and z represents: ##STR31##
Claim 7
Dependent Claim
44 words
A complex in accordance with claim 3 wherein said detectable polypeptide is linked to an indicator molecule selected from the group consisting of fluorescent dyes, electron-dense reagents, enzymes which can be reacted with a substrate to produce a visually detectable reaction product, and radioisotopes.
Claim 8
Dependent Claim
34 words
A complex in accordance with claim 4 wherein said detectable polypeptide is a fluorescent dye, electron dense reagent, or enzyme which can be reacted with a substrate to produce a visually detectable reaction product.
Claim 9
Dependent Claim
252 words
A complex useful as a probe for detecting the presence or absence of a nucleic acid, said complex comprising a detectable polypeptide complexed with an oligo- or polynucleotide containing a nucleotide having the structure: ##STR32## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety covalently bonded to the C 1 '-position of the sugar moiety, provided that whenever B is purine or 7-deazapurine, the sugar moiety is attached at the N 9 -position of the purine or deazapurine, and whenever B is a pyrimidine, the sugar moiety is attached at the N 1 -position of the pyrimidine; wherein A represents at least three carbon atoms, is capable of specifically complexing with the detectable polypeptide when A is linked to B, and represents a component of a signalling moiety capable of producing a detectable signal; wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with the characteristic ability of A to form said complex with the detectable polypeptide; wherein if B is a purine, A or the linkage group is attached to the 8-position of the purine, if B is a 7-deazapurine, A or the linkage group is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A or the linkage group is attached to the 5-position of the pyrimidine; wherein one of x and y represents ##STR33## and the other of x and y is absent or represents --OH or --H; and wherein z represents H-- or HO--.
Claim 10
Dependent Claim
252 words
A complex useful as a probe for detecting the presence or absence of a nucleic acid, said complex comprising a detectable polypeptide complexed with an oligo- or polynucleotide containing a nucleotide having the structure: ##STR32## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety covalently bonded to the C 1 '-position of the sugar moiety, provided that whenever B is purine or 7-deazapurine, the sugar moiety is attached at the N 9 -position of the purine or deazapurine, and whenever B is a pyrimidine, the sugar moiety is attached at the N 1 -position of the pyrimidine; wherein A represents at least three carbon atoms, is capable of specifically complexing with the detectable polypeptide when A is linked to B, and represents a component of a signalling moiety capable of producing a detectable signal; wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with the characteristic ability of A to form said complex with the detectable polypeptide; wherein if B is a purine, A or the linkage group is attached to the 8-position of the purine, if B is a 7-deazapurine, A or the linkage group is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A or the linkage group is attached to the 5-position of the pyrimidine; wherein one of x and y represents ##STR33## and the other of x and y is absent or represents --OH or --H; and wherein z represents H-- or HO--.
Claim 11
Dependent Claim
44 words
A complex in accordance with claim 6 wherein said detectable polypeptide is linked to an indicator molecule selected from the group consisting of fluorescent dyes, electron-dense reagents, enzymes which can be reacted with a substrate to produce a visually detectable reaction product, and radioisotopes.
Claim 12
Dependent Claim
34 words
A complex in accordance with claim 7 wherein said detectable polypeptide is a fluorescent dye, electron dense reagent, or enzyme which can be reacted with a substrate to produce a visually detectable reaction product.
Claim 13
Dependent Claim
14 words
A complex in accordance with claims 3 or 6 wherein A is a ligand.
Claim 14
Dependent Claim
21 words
A complex according to claim 9 wherein said ligand is selected from the group consisting of biotin, iminobiotin, or a cofactor.
Claim 15
Dependent Claim
21 words
A complex in accordance with claim 9 wherein said ligand is selected from the group consisting of antigens, antibodies and haptens.
Claim 16
Dependent Claim
12 words
A complex in accordance with claim 11 wherein said ligand is dinitrophenol.
Claim 17
Dependent Claim
58 words
A complex according to claims 3 or 6 wherein the detectable polypeptide is indirectly detectable by specifically complexing the detectable polypeptide with a second moiety covalently linked to an indicator molecule selected from the group consisting of fluorescent dyes, electron-dense reagents, enzymes which can be reacted with a substrate to produce a visually detectable reaction product, and radioisotopes.
Claim 18
Dependent Claim
34 words
A complex according to claim 13 wherein the detectable polypeptide is selected from the group consisting of avidin and streptavidin and the second moiety is selected from the group consisting of biotin and iminobiotin.
Claim 19
Dependent Claim
27 words
A complex according to claim 14 wherein the indicator molecule is an enzyme which can be reacted with a substrate to produce a visually detectable reaction product.
Claim 20
Dependent Claim
17 words
The complex of claims 3 or 6 wherein said linkage group is comprises a --CH 2 --NH--.
Claim 21
Dependent Claim
29 words
The complex of claims 3 or 6 wherein said linkage group is selected from the group consisting of --CH═CH--CH 2 --NH-- and --CH═CH--CH 2 --O--CH 2 --CH(OH)--CH 2 --NH--.
Claim 22
Dependent Claim
17 words
The complex of claims 3 or 6 wherein A is an allylamine group linked directly to B.
Claim 23
Dependent Claim
16 words
The complex of claims 3 or 6 wherein A is a moiety comprising an olefinic bond.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1998-12-18 | 1999-06-19 | 1999-12-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2002-12-18 | 2003-06-19 | 2003-12-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2006-12-18 | 2007-06-19 | 2007-12-19 | $7700.00 | Window Closed / Lapsed |