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US5780269A
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Hybrid molecules
Assignee: Rutgers Health
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Filing: 1995-07-18
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Granted: 1998-07-14
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Expires: 2015-07-18
Abstract
Abstract Novel hybrid molecules comprising an RNA and a DNA portion are disclosed. A method for synthesizing the molecule is disclosed.
Inventors
- 👤 Sumiko Inouye
- 👤 Masayori Inouye
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 04:53
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/10; C12N9/12; C12N9/1241; C12N9/1276; C12N15/00; C12N15/09; C12N15/10; C12P; C12P19/00; C12P19/26; C12P19/28; C12P19/30; C12P19/34
Claims Analysis
Total Claims: 34
Claim 1
Independent Claim
121 words
A molecule which has features as shown below ##STR2## wherein X is 5 to 11 nucleotides in length, Y is 7 to 69 nucleotides in length, Z is 3 to 19 nucleotides in length, L is 9 to 26 nucleotides in length, S is 34 to 136 nucleotides in length, and the total nucleotide length of W, V, and Q is between 114 and 239 minus the sum of X, Y, Z, L, and S, wherein the DNA portion of the molecule has one stem-and-loop structure and the RNA portion of the molecule has at least one stem-and-loop structure, wherein said lengths are determined in number of nucleotides, which molecule has in its RNA portion a foreign DNA or RNA sequence.
Claim 2
Dependent Claim
121 words
A molecule which has features as shown below ##STR2## wherein X is 5 to 11 nucleotides in length, Y is 7 to 69 nucleotides in length, Z is 3 to 19 nucleotides in length, L is 9 to 26 nucleotides in length, S is 34 to 136 nucleotides in length, and the total nucleotide length of W, V, and Q is between 114 and 239 minus the sum of X, Y, Z, L, and S, wherein the DNA portion of the molecule has one stem-and-loop structure and the RNA portion of the molecule has at least one stem-and-loop structure, wherein said lengths are determined in number of nucleotides, which molecule has in its RNA portion a foreign DNA or RNA sequence.
Claim 3
Dependent Claim
15 words
The molecule of claim 1 wherein there are two stem-loop structures in the RNA portion.
Claim 4
Independent Claim
137 words
A method for synthesizing a hybrid molecule which molecule comprises a single-stranded RNA covalently linked to a single-stranded DNA by a 2',5'-phosphodiester bond between the 2'-OH group of an internal rG residue and the 5'-phosphate of the DNA strand and non-covalently linked to the DNA by overlapping complementary nucleotides at the 3' ends of the RNA and DNA strands, which RNA and DNA portions each form a secondary structure, which method comprises: a) transforming a bacterium or a yeast with a plasmid containing a gene for reverse transcriptase and a template for the reverse transcriptase (RT), which template has msr and msd genes, which msr and msd genes are annealed at their 3' ends to each other, b) allowing the bacterium or yeast to synthesize the hybrid molecule, and c) isolating the synthetic hybrid molecule so produced.
Claim 5
Independent Claim
137 words
A method for synthesizing a hybrid molecule which molecule comprises a single-stranded RNA covalently linked to a single-stranded DNA by a 2',5'-phosphodiester bond between the 2'-OH group of an internal rG residue and the 5'-phosphate of the DNA strand and non-covalently linked to the DNA by overlapping complementary nucleotides at the 3' ends of the RNA and DNA strands, which RNA and DNA portions each form a secondary structure, which method comprises: a) transforming a bacterium or a yeast with a plasmid containing a gene for reverse transcriptase and a template for the reverse transcriptase (RT), which template has msr and msd genes, which msr and msd genes are annealed at their 3' ends to each other, b) allowing the bacterium or yeast to synthesize the hybrid molecule, and c) isolating the synthetic hybrid molecule so produced.
Claim 6
Dependent Claim
39 words
The method of claim 3 which, prior to synthesis of the hybrid molecule, comprising the msr and msd genes from oligonucleotides, and annealing the 3' ends of the genes to each other, thus obtaining a template for the RT.
Claim 7
Dependent Claim
17 words
The method of claim 3 wherein the RT gene is under the control of an inducible promoter.
Claim 8
Dependent Claim
17 words
The method of claim 3 wherein the template is upstream of the RT gene in the plasmid.
Claim 9
Independent Claim
149 words
A method for synthesizing a hybrid molecule which molecule comprises a single-stranded RNA covalently linked to a single-stranded DNA by a 2',5'-phosphodiester bond between the 2'-OH group of an internal rG residue and the 5'-phosphate of the DNA strand and non-covalently linked to the DNA by overlapping complementary nucleotides at the 3' ends of the RNA and DNA strands, which RNA and DNA portions each form a secondary structure, which method comprises: a) synthesizing an msr gene and an msd gene from oligonucleotides, b) annealing the msr and msd genes at their respective 3' ends, thereby obtaining a template for a reverse transcriptase, d) inserting the template and a gene for a bacterial reverse transcriptase (RT) into a plasmid, c) transforming a bacterium or a yeast with the plasmid, d) allowing the bacterium or yeast to produce the hybrid molecule, and e) isolating the synthetic hybrid molecule so produced.
Claim 10
Independent Claim
149 words
A method for synthesizing a hybrid molecule which molecule comprises a single-stranded RNA covalently linked to a single-stranded DNA by a 2',5'-phosphodiester bond between the 2'-OH group of an internal rG residue and the 5'-phosphate of the DNA strand and non-covalently linked to the DNA by overlapping complementary nucleotides at the 3' ends of the RNA and DNA strands, which RNA and DNA portions each form a secondary structure, which method comprises: a) synthesizing an msr gene and an msd gene from oligonucleotides, b) annealing the msr and msd genes at their respective 3' ends, thereby obtaining a template for a reverse transcriptase, d) inserting the template and a gene for a bacterial reverse transcriptase (RT) into a plasmid, c) transforming a bacterium or a yeast with the plasmid, d) allowing the bacterium or yeast to produce the hybrid molecule, and e) isolating the synthetic hybrid molecule so produced.
Claim 11
Dependent Claim
82 words
An isolated hybrid molecule which comprises a single-stranded RNA portion covalently linked to a single-stranded DNA portion by a 2',5'-phosphodiester bond between the 2'-OH group of an internal rG residue and the 5'-phosphate of the DNA portion and non-covalently linked to the DNA portion by overlapping complementary nucleotides at the 3' ends of the RNA and DNA portions, which RNA and DNA portions each form a stem-loop structure, wherein the number of nucleotides in the DNA portion is between 67 and 162.
Claim 12
Dependent Claim
82 words
An isolated hybrid molecule which comprises a single-stranded RNA portion covalently linked to a single-stranded DNA portion by a 2',5'-phosphodiester bond between the 2'-OH group of an internal rG residue and the 5'-phosphate of the DNA portion and non-covalently linked to the DNA portion by overlapping complementary nucleotides at the 3' ends of the RNA and DNA portions, which RNA and DNA portions each form a stem-loop structure, wherein the number of nucleotides in the DNA portion is between 67 and 162.
Claim 13
Dependent Claim
26 words
The hybrid molecule of claim 8 wherein there is one stem-loop structure in the DNA portion and at least one stem-loop structures in the RNA portion.
Claim 14
Dependent Claim
14 words
The hybrid molecule of claim 8 wherein the RNA portion has two stem-loop structures.
Claim 15
Dependent Claim
20 words
The hybrid molecule of claim 8 wherein the number of nucleotides in the RNA portion is between 49 and 82.
Claim 16
Dependent Claim
28 words
The hybrid molecule of claim 8 wherein the number of overlapping complementary nucleotides at the 3' ends of the RNA and DNA portions is between 5 and 11.
Claim 17
Dependent Claim
34 words
The hybrid molecule of claim 8 which has in its DNA or RNA portion a foreign DNA or RNA sequence which is an antisense sequence with respect to an mRNA of a target protein.
Claim 18
Dependent Claim
25 words
The molecule of claim 1 wherein the foreign DNA or RNA sequence is an antisense molecule with respect to an mRNA of a target protein.
Claim 19
Dependent Claim
80 words
A hybrid molecule which comprises a single-stranded RNA portion covalently linked to a single-stranded DNA portion by a 2',5'-phosphodiester bond between the 2'-OH group of an internal rG residue and the 5'-phosphate of the DNA portion and non-covalently linked to the DNA portion by overlapping complementary nucleotides at the 3' ends of the RNA and DNA portions, which RNA and DNA portions each form a stem-loop structure, and a foreign DNA or RNA sequence in the DNA or RNA portion.
Claim 20
Dependent Claim
80 words
A hybrid molecule which comprises a single-stranded RNA portion covalently linked to a single-stranded DNA portion by a 2',5'-phosphodiester bond between the 2'-OH group of an internal rG residue and the 5'-phosphate of the DNA portion and non-covalently linked to the DNA portion by overlapping complementary nucleotides at the 3' ends of the RNA and DNA portions, which RNA and DNA portions each form a stem-loop structure, and a foreign DNA or RNA sequence in the DNA or RNA portion.
Claim 21
Dependent Claim
26 words
The hybrid molecule of claim 15 wherein there is one stem-loop structure in the DNA portion and at least one stem-loop structures in the RNA portion.
Claim 22
Dependent Claim
14 words
The hybrid molecule of claim 15 wherein the RNA portion has two stem-loop structures.
Claim 23
Dependent Claim
20 words
The hybrid molecule of claim 15 wherein the number of nucleotides in the DNA portion is between 65 and 163.
Claim 24
Dependent Claim
20 words
The hybrid molecule of claim 15 wherein the number of nucleotides in the RNA portion is between 49 and 82.
Claim 25
Dependent Claim
28 words
The hybrid molecule of claim 15 wherein the number of overlapping complementary nucleotides at the 3' ends of the RNA and DNA portions is between 5 and 11.
Claim 26
Dependent Claim
9 words
The hybrid molecule of claim 15 which is isolated.
Claim 27
Dependent Claim
99 words
A hybrid molecule which comprises a single-stranded RNA portion covalently linked to a single-stranded DNA portion by a 2',5'-phosphodiester bond between the 2'-OH group of an internal rG residue and the 5'-phosphate of the DNA portion and non-covalently linked to the DNA portion by overlapping complementary nucleotides at the 3' ends of the RNA and DNA portions, which RNA and DNA portions each form a stem-loop structure, and a foreign DNA or RNA sequence in the DNA or RNA portion, which foreign DNA or RNA sequence is an antisense sequence with respect to an mRNA of a target protein.
Claim 28
Dependent Claim
99 words
A hybrid molecule which comprises a single-stranded RNA portion covalently linked to a single-stranded DNA portion by a 2',5'-phosphodiester bond between the 2'-OH group of an internal rG residue and the 5'-phosphate of the DNA portion and non-covalently linked to the DNA portion by overlapping complementary nucleotides at the 3' ends of the RNA and DNA portions, which RNA and DNA portions each form a stem-loop structure, and a foreign DNA or RNA sequence in the DNA or RNA portion, which foreign DNA or RNA sequence is an antisense sequence with respect to an mRNA of a target protein.
Claim 29
Dependent Claim
26 words
The hybrid molecule of claim 22 wherein there is one stem-loop structure in the DNA portion and at least one stem-loop structures in the RNA portion.
Claim 30
Dependent Claim
14 words
The hybrid molecule of claim 22 wherein the RNA portion has two stem-loop structures.
Claim 31
Dependent Claim
20 words
The hybrid molecule of claim 22 wherein the number of nucleotides in the DNA portion is between 65 and 163.
Claim 32
Dependent Claim
20 words
The hybrid molecule of claim 22 wherein the number of nucleotides in the RNA portion is between 49 and 82.
Claim 33
Dependent Claim
28 words
The hybrid molecule of claim 22 wherein the number of overlapping complementary nucleotides at the 3' ends of the RNA and DNA portions is between 5 and 11.
Claim 34
Dependent Claim
9 words
The hybrid molecule of claim 22 which is isolated.
References Cited (Backward Citations)
US5405775A
Cited
US5436141A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2001-07-13 | 2002-01-12 | 2002-07-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2005-07-13 | 2006-01-12 | 2006-07-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2009-07-13 | 2010-01-12 | 2010-07-14 | $7700.00 | Window Closed / Lapsed |