US5912340A
Expired / Lapsed
US
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Selective binding complementary oligonucleotides
Assignee: Epoch Biosciences Inc
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Filing: 1995-10-04
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Granted: 1999-06-15
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Expires: 2015-10-04
Abstract
Abstract In a matched pair of oligonucleotides (ODNS) each member of the pair is complementary or substantially complementary in the Watson Crick sense to a target sequence of duplex nucleic acid where the two strands of the target sequence are themselves complementary to one another. The ODNs include modified bases of such nature that the modified base forms a stable hydrogen bonded base pair with the natural partner base, but does not form a stable hydrogen bonded base pair with its modified partner. This is accomplished when in a hybridized structure the modified base is capable of forming two or more hydrogen bonds with its natural complementary base, but only one hydrogen bond with its modified partner. Due to the lack of stable hydrogen bonding with each other, the matched pair of oligonucleotides have a melting temperature under physiological or substantially physiological conditions of approximately 40 DEG C. or less. However each of the matched ODN pair of the invention forms a substantially stable hybrid with the target sequence in each strand of the duplex nucleic acid. The hybrids of target duplex nucleic acids formed with the ODN pairs of the invention are useful for gene mapping and in diagnostic and therapeutic applications.
Inventors
- 👤 Igor V. Kutyavin
- 👤 Jinsuk Woo
- 👤 Eugeny A. Lukhtanov
- 👤 Rich B. Meyer, Jr.
- 👤 Howard B. Gamper
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 17:53
CPC Classifications
C; C07; C07H; C07H21/00; C12; C12Q; C12Q1/00; C12Q1/68
Claims Analysis
Total Claims: 27
Claim 1
Independent Claim
205 words
A pair of oligonucleotides (ODNs), each of said ODNs comprising nucleotide moieties having naturally occurring aglycon bases and a combination of modified aglycon bases selected from the group consisting of the combinations (1) A', T', (2) G', C' and (3) A', T', G', C', the duplex form of said pair of ODNs having a melting temperature under physiological conditions of less than approximately 40° C., each of said pair of ODNs being substantially complementary in the Watson-Crick sense to one of the two strands of a duplexed target sequence in nucleic acid, wherein the nucleotide moieties having the modified bases have the following properties: within complementary oligonucleotides A' does not form a stable hydrogen bonded base pair with T' and forms a stable hydrogen bonded base pair with T; within complementary oligonucleotides T' does not form a stable hydrogen bonded base pair with A' and forms a stable hydrogen bonded base pair with A; within complementary oligonucleotides G' does not form a stable hydrogen bonded base pair with C' and forms a stable hydrogen bonded base pair with C, and within complementary oligonucleotides C' does not form a stable hydrogen bonded base pair with GI and forms a stable hydrogen bonded base pair with G.
Claim 2
Dependent Claim
205 words
A pair of oligonucleotides (ODNs), each of said ODNs comprising nucleotide moieties having naturally occurring aglycon bases and a combination of modified aglycon bases selected from the group consisting of the combinations (1) A', T', (2) G', C' and (3) A', T', G', C', the duplex form of said pair of ODNs having a melting temperature under physiological conditions of less than approximately 40° C., each of said pair of ODNs being substantially complementary in the Watson-Crick sense to one of the two strands of a duplexed target sequence in nucleic acid, wherein the nucleotide moieties having the modified bases have the following properties: within complementary oligonucleotides A' does not form a stable hydrogen bonded base pair with T' and forms a stable hydrogen bonded base pair with T; within complementary oligonucleotides T' does not form a stable hydrogen bonded base pair with A' and forms a stable hydrogen bonded base pair with A; within complementary oligonucleotides G' does not form a stable hydrogen bonded base pair with C' and forms a stable hydrogen bonded base pair with C, and within complementary oligonucleotides C' does not form a stable hydrogen bonded base pair with GI and forms a stable hydrogen bonded base pair with G.
Claim 3
Dependent Claim
99 words
The ODNs of claim 1 wherein the nucleotide moiety A' has the structure selected from the groups shown by formulas (i), (ii) and (iii) ##STR10## wherein X is N or CH; Y is O or S; Z is OH or CH 3 ; R is H, F, or OR 2 , where R 2 is H, C 1-6 alkyl or allyl, and R 1 is C 1-4 alkyl, C 1-4 alkoxy, C 1-4 alkylthio, F, or NHR 3 where R 3 is H, or C 1-4 alkyl, and where R* is H, a cross-linking function or a reporter group.
Claim 4
Dependent Claim
67 words
The ODNs of claim 1 wherein the nucleotide moiety T' has the formula (iv) ##STR11## wherein Y is O or S; Z is OH or CH 3 ; R is H, F, or OR 2 , where R 2 is H, C 1-6 alkyl or allyl, and R 4 is H, C 1-6 alkyl, C 1-6 alkenyl, C 1-6 alkynyl, a cross-linking function or a reporter group.
Claim 5
Dependent Claim
100 words
The ODNs of claim 1 wherein the nucleotide moiety G' has the structure selected from the groups shown by formulas (v), (vi) and (vii) ##STR12## wherein X is N or CH; Y is O or S; Z is OH or CH 3 ; R is H, F, or OR 2 , where R 2 is H, C 1-6 alkyl or allyl, and R 1 is H, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 alkylthio, F, or NHR 3 where R 3 is H, or C 1-4 alkyl, and where R* is H, a cross-linking function or a reporter group.
Claim 6
Dependent Claim
89 words
The ODNs of claim 1 wherein the nucleotide C' has the structure selected from the groups shown by formulas (viii) and (ix) ##STR13## wherein Y is O or S; Z is OH or CH 3 ; R is H, F, or OR 2 , where R 2 is H, C 1-6 alkyl or allyl, R 4 is H, C 1-6 alkyl, C 1-6 alkenyl, C 1-6 alknyl, a cross-linking function or a reporter group; Z 1 is O or NH, and R 5 is H, or C 1-4 alkyl.
Claim 7
Dependent Claim
18 words
The ODNs of claim 2 wherein the nucleotide moiety A' has the structure in accordance with formula (i).
Claim 8
Dependent Claim
16 words
The ODNs of claim 6 wherein X is N, Z is OH, and Y is O.
Claim 9
Dependent Claim
12 words
The ODNs of claim 7 wherein R 1 is NH 2 .
Claim 10
Dependent Claim
13 words
The ODNs of claim 3 wherein Z is OH, and Y is O.
Claim 11
Dependent Claim
12 words
The ODNs of claim 9 wherein R 4 is CH 3 .
Claim 12
Dependent Claim
18 words
The ODNs of claim 4 wherein the nucleotide moiety G' has the structure in accordance with formula (v).
Claim 13
Dependent Claim
16 words
The ODNs of claim 11 wherein X is N, Z is OH, and Y is O.
Claim 14
Dependent Claim
10 words
The ODNs of claim 12 wherein R 1 is H.
Claim 15
Dependent Claim
18 words
The ODNs of claim 5 wherein the nucleotide moiety C' has the structure in accordance with formula (viii).
Claim 16
Dependent Claim
17 words
The ODNs of claim 14 wherein, Z is OH, Z 1 is NH and Y is O.
Claim 17
Dependent Claim
10 words
The ODNs of claim 15 wherein R 5 is H.
Claim 18
Dependent Claim
12 words
The ODNs of claim 1 having approximately 5 to 99 nucleotide units.
Claim 19
Dependent Claim
13 words
The ODNs of claim 1 wherein each of the nucleotides is a 2'-deoxyribonucleotide.
Claim 20
Dependent Claim
13 words
The ODNs of claim 1 wherein each of the nucleotides is a ribonucleotide.
Claim 21
Dependent Claim
15 words
The ODNs of claim 1 comprising at least one nucleotide unit having a 2-O-methylribose moiety.
Claim 22
Dependent Claim
17 words
The ODNs of claim 1 comprising a cross-linking function covalently attached to at least one nucleotide unit.
Claim 23
Dependent Claim
9 words
The ODNs of claim 1 comprising a reporter group.
Claim 24
Dependent Claim
15 words
The ODNs of claim 1 wherein the combination of modified aglycon bases is A', T'.
Claim 25
Dependent Claim
15 words
The ODNs of claim 1 wherein the combination of modified aglycon bases is G', C'.
Claim 26
Dependent Claim
17 words
The ODNs of claim 1 wherein the combination of modified aglycon bases is A', T', G', C'.
Claim 27
Dependent Claim
20 words
The ODNs of claim 1 where the pair of oligonucleotides are linked to one another by a covalently bonded tether.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2002-06-14 | 2002-12-14 | 2003-06-15 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2006-06-14 | 2006-12-14 | 2007-06-15 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2010-06-14 | 2010-12-14 | 2011-06-15 | $7700.00 | Window Closed / Lapsed |