US5208149A
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Nucleic acid constructs containing stable stem and loop structures
Assignee: Research Foundation of the State University of New York
•
Filing: 1992-04-10
•
Granted: 1993-05-04
•
Expires: 2012-04-10
Abstract
Abstract Gene expression in a cell can be regulated or inhibited by incorporating into or associating with the genetic material of the cell a non-native nucleic acid sequence which is transcribed to produce an mRNA which is complementary to and capable of binding to the mRNA produced by the genetic material of said cell.
Inventors
- 👤 Masayori Inouye
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 20:35
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/113; C07; C07H; C07H21/00; A; A61; A61K; A61K38/00; C12N2310/00; C12N2310/10; C12N2310/11; C12N2310/111
Claims Analysis
Total Claims: 204
Claim 1
Independent Claim
74 words
A non-native polynucleotide construct comprising: a. a transcriptional promoter segment; b. a segment coding for a stable stem and loop structure with a negative ΔG of formation operably linked downstream of said promoter segment; and c. a polynucleotide segment comprising a gene segment operably linked downstream of said promoter segment and inverted with respect to a gene in a cell, whereby the transcript of said inverted gene segment regulates the function of said gene.
Claim 2
Dependent Claim
74 words
A non-native polynucleotide construct comprising: a. a transcriptional promoter segment; b. a segment coding for a stable stem and loop structure with a negative ΔG of formation operably linked downstream of said promoter segment; and c. a polynucleotide segment comprising a gene segment operably linked downstream of said promoter segment and inverted with respect to a gene in a cell, whereby the transcript of said inverted gene segment regulates the function of said gene.
Claim 3
Dependent Claim
25 words
The construct of claim 1 wherein the stem and loop structure is at the 3' end of the transcript produced from said inverted gene segment.
Claim 4
Dependent Claim
25 words
The construct of claim 1 wherein the stem and loop structure is at the 5' end of the transcript produced from said inverted gene segment.
Claim 5
Dependent Claim
15 words
The construct of claim 1 wherein the ΔG of formation is at least -4.5 Kcal/mol.
Claim 6
Dependent Claim
15 words
The construct of claim 1 wherein the ΔG of formation is at least -12.5 Kcal/mol.
Claim 7
Dependent Claim
17 words
The construct of claim 1 wherein the stem and loop structure is derived from a prokaryotic RNA.
Claim 8
Dependent Claim
17 words
The construct of claim 1 wherein the stem and loop structure is derived from a eukaryotic RNA.
Claim 9
Dependent Claim
34 words
The construct of claim 1 wherein the stem and loop structure is derived from an RNA selected from the group consisting of tRNA, mRNA, 5S RNA, rRNA, hnRNA, viroid RNA and viral genomic ssRNA.
Claim 10
Dependent Claim
29 words
The construct of claim 1 wherein the stem and loop structure is derived from an RNA selected from the group consisting of tRNA, mRNA, 5S RNA, rRNA and hnRNA.
Claim 11
Dependent Claim
16 words
The construct of claim 1 wherein the stem and loop structure is derived from a rRNA.
Claim 12
Dependent Claim
16 words
The construct of claim 1 wherein the stem and loop structure is derived from a tRNA.
Claim 13
Dependent Claim
17 words
The construct of claim 1 wherein the stem and loop structure is derived from a mRNA sequence.
Claim 14
Dependent Claim
19 words
The construct of claim 1 wherein said transcriptional promoter segment of said non-native polynucleotide construct comprises an inducible promoter.
Claim 15
Dependent Claim
11 words
The construct of claim 1 wherein said gene is an oncogene.
Claim 16
Dependent Claim
12 words
The construct of claim 1 wherein said gene is a viral gene.
Claim 17
Dependent Claim
21 words
The construct of claim 1 wherein said gene encodes a positive regulatorwhose presence is necessary for the expression of another gene.
Claim 18
Dependent Claim
16 words
The construct of claim 1 wherein said gene is related to a genetic disease or defect.
Claim 19
Dependent Claim
11 words
The construct of claim 1 wherein said gene encodes a protein.
Claim 20
Dependent Claim
30 words
The construct of claim 18 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the coding region of the RNA transcript produced by said gene.
Claim 21
Dependent Claim
30 words
The construct of claim 18 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the non-coding region of the RNA transcript produced by said gene.
Claim 22
Dependent Claim
31 words
The construct of claim 18 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the 5' non-coding region of the RNA transcript produced by said gene.
Claim 23
Dependent Claim
31 words
The construct of claim 18 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the ribosome binding region of the RNA transcript produced by said gene.
Claim 24
Dependent Claim
31 words
The construct of claim 18 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the translation initiation region of the RNA transcript produced by said gene.
Claim 25
Dependent Claim
36 words
The construct of claim 18 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the ribosome binding region and the translation initiation region of the RNA transcript produced by said gene.
Claim 26
Dependent Claim
15 words
The construct of claim 1 wherein said non-native polynucleotide construct is incorporated into a vector.
Claim 27
Dependent Claim
11 words
The construct of claim 25 wherein said vector is a plasmid.
Claim 28
Dependent Claim
12 words
The construct of claim 25 wherein said vector is a viral vector.
Claim 29
Dependent Claim
12 words
The construct of claim 25 wherein said vector is either single-strandedor double-stranded.
Claim 30
Dependent Claim
19 words
The construct of any one of claims 1 to 28 wherein said non-native polynucleotide construct is a DNA construct.
Claim 31
Dependent Claim
16 words
A pharmaceutical composition which comprises the polynucleotide construct of any one of claims 1 to 28.
Claim 32
Dependent Claim
57 words
A non-native polynucleotide construct which produces in a cell, a non-naturally occurring polynucleotide complementary to a RNA transcript produced by a gene in said cell, said non-naturally occurring polynucleotide further comprising a stable stem and loop structure with a negative ΔG of formation, whereby said non-naturally occurring polynucleotide regulates the function of said gene in said cell.
Claim 33
Dependent Claim
57 words
A non-native polynucleotide construct which produces in a cell, a non-naturally occurring polynucleotide complementary to a RNA transcript produced by a gene in said cell, said non-naturally occurring polynucleotide further comprising a stable stem and loop structure with a negative ΔG of formation, whereby said non-naturally occurring polynucleotide regulates the function of said gene in said cell.
Claim 34
Dependent Claim
21 words
The construct of claim 31 wherein the stem and loop structure is at the 3' end of the non-naturally occurring polynucleotide.
Claim 35
Dependent Claim
21 words
The construct of claim 31 wherein the stem and loop structure is at the 5' end of the non-naturally occurring polynucleotide.
Claim 36
Dependent Claim
15 words
The construct of claim 31 wherein the ΔG of formation is at least -4.5 Kcal/mol.
Claim 37
Dependent Claim
15 words
The construct of claim 31 wherein the ΔG of formation is at least -12.5 Kcal/mol.
Claim 38
Dependent Claim
17 words
The construct of claim 31 wherein the stem and loop structure is derived from a prokaryotic RNA.
Claim 39
Dependent Claim
17 words
The construct of claim 31 wherein the stem and loop structure is derived from a eukaryotic RNA.
Claim 40
Dependent Claim
34 words
The construct of claim 31 wherein the stem and loop structure is derived from an RNA selected from the group consisting of tRNA, mRNA, 5S RNA, rRNA, hRNA, viroid RNA and viral genomic ssRNA.
Claim 41
Dependent Claim
29 words
The construct of claim 31 wherein the stem and loop structure is derived from an RNA selected from the group consisting of tRNA, mRNA, 5S RNA, rRNA and hnRNA.
Claim 42
Dependent Claim
16 words
The construct of claim 31 wherein the stem and loop structure is derived from a rRNA.
Claim 43
Dependent Claim
16 words
The construct of claim 31 wherein the stem and loop structure is derived from a tRNA.
Claim 44
Dependent Claim
16 words
The construct of claim 31 wherein the stem and loop structure is derived from a mRNA.
Claim 45
Dependent Claim
12 words
The construct of claim 31 wherein said construct comprises an inducible promoter.
Claim 46
Dependent Claim
11 words
The construct of claim 31 wherein said gene is an oncogene.
Claim 47
Dependent Claim
12 words
The construct of claim 31 wherein said gene is a viral gene.
Claim 48
Dependent Claim
22 words
The construct of claim 31 wherein said gene encodes a positive regulator whose presence is necessary for the expression of another gene.
Claim 49
Dependent Claim
16 words
The construct of claim 31 wherein said gene is related to a genetic disease or defect.
Claim 50
Dependent Claim
11 words
The construct of claim 31 wherein said gene encodes a protein.
Claim 51
Dependent Claim
28 words
The construct of claim 48 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the coding region of the RNA transcript produced by said gene.
Claim 52
Dependent Claim
28 words
The construct of claim 49 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the non-coding region of the RNA transcript produced by said gene.
Claim 53
Dependent Claim
29 words
The construct of claim 49 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the 5' non-coding region of the RNA transcript produced by said gene.
Claim 54
Dependent Claim
29 words
The construct of claim 49 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the ribosome binding region of the RNA transcript produced by said gene.
Claim 55
Dependent Claim
29 words
The construct of claim 49 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the translation initiation region of the RNA transcript produced by said gene.
Claim 56
Dependent Claim
34 words
The construct of claim 49 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the ribosome binding region and the translation initiation region of said RNA transcript produced by said gene.
Claim 57
Dependent Claim
15 words
The construct of claim 49 wherein the non-native polynucleotide construct is incorporated into a vector.
Claim 58
Dependent Claim
11 words
The construct of claim 55 wherein said vector is a plasmid.
Claim 59
Dependent Claim
12 words
The construct of claim 55 wherein said vector is a viral vector.
Claim 60
Dependent Claim
13 words
The construct of claim 55 wherein said vector is either single-stranded or double-stranded.
Claim 61
Dependent Claim
16 words
The construct any one of claim 31 and 58 wherein said non-native polynucleotide construct is DNA.
Claim 62
Dependent Claim
16 words
A pharmaceutical composition which comprises the polynucleotide construct of any one of claims 31 to 58.
Claim 63
Dependent Claim
77 words
A cell comprising a non-native polynucleotide construct comprising: (a) a transcriptional promoter segment; (b) a segment coding for a stable stem and loop structure with a negative ΔG of formation operably linked downstream of said promoter segment; and (c) a polynucleotide segment comprising a gene segment operably linked downstream of said promoter segment and inverted with respect to a gene in a cell, whereby the transcript of said inverted gene segment regulates the function of said gene.
Claim 64
Dependent Claim
77 words
A cell comprising a non-native polynucleotide construct comprising: (a) a transcriptional promoter segment; (b) a segment coding for a stable stem and loop structure with a negative ΔG of formation operably linked downstream of said promoter segment; and (c) a polynucleotide segment comprising a gene segment operably linked downstream of said promoter segment and inverted with respect to a gene in a cell, whereby the transcript of said inverted gene segment regulates the function of said gene.
Claim 65
Dependent Claim
25 words
The cell of claim 61 wherein the stem and loop structure is at the 3' end of the transcript produced from said inverted gene segment.
Claim 66
Dependent Claim
25 words
The cell of claim 61 wherein the stem and loop structure is at the 5' end of the transcript produced from said inverted gene segment.
Claim 67
Dependent Claim
15 words
The cell of claim 61 wherein the ΔG of formation is at least -4.5 Kcal/mol.
Claim 68
Dependent Claim
15 words
The cell of claim 61 wherein the ΔG of formation is at least -12.5 Kcal/mol.
Claim 69
Dependent Claim
17 words
The cell of claim 61 wherein the stem and loop structure is derived from a prokaryotic RNA.
Claim 70
Dependent Claim
17 words
The cell of claim 61 wherein the stem and loop structure is derived from a eukaryotic RNA.
Claim 71
Dependent Claim
34 words
The cell of claim 61 wherein the stem and loop structure is derived from an RNA selected from the group consisting of tRNA, mRNA, 5S RNA, rRNA, hnRNA, viroid RNA and viral genomic ssRNA.
Claim 72
Dependent Claim
29 words
The cell of claim 61 wherein the stem and loop structure is derived from an RNA selected from the group consisting of tRNA, mRNA, 5S RNA, rRNA and hnRNA.
Claim 73
Dependent Claim
16 words
The cell of claim 61 wherein the stem and loop structure is derived from a rRNA.
Claim 74
Dependent Claim
16 words
The cell of claim 61 wherein the stem and loop structure is derived from a tRNA.
Claim 75
Dependent Claim
16 words
The cell of claim 61 wherein the stem and loop structure is derived from a mRNA.
Claim 76
Dependent Claim
19 words
The cell of claim 61 wherein said transcriptional promoter segment of said non-native polynucleotide construct comprises an inducible promoter.
Claim 77
Dependent Claim
11 words
The cell of claim 61 wherein said gene is an oncogene.
Claim 78
Dependent Claim
12 words
The cell of claim 61 wherein said gene is a viral gene.
Claim 79
Dependent Claim
22 words
The cell of claim 61 wherein said gene encodes a positive regulator whose presence is necessary for the expression of another gene.
Claim 80
Dependent Claim
16 words
The cell of claim 61 wherein said gene is related to a genetic disease or defect.
Claim 81
Dependent Claim
11 words
The cell of claim 61 wherein said gene encodes a protein.
Claim 82
Dependent Claim
30 words
The cell of claim 78 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the coding region of the RNA transcript produced by said gene.
Claim 83
Dependent Claim
30 words
The cell of claim 78 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the non-coding region of the RNA transcript produced by said gene.
Claim 84
Dependent Claim
31 words
The cell of claim 78 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the 5' non-coding region of the RNA transcript produced by said gene.
Claim 85
Dependent Claim
31 words
The cell of claim 78 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the ribosome binding region of the RNA transcript produced by said gene.
Claim 86
Dependent Claim
31 words
The cell of claim 78 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the translation initiation region of the RNA transcript produced by said gene.
Claim 87
Dependent Claim
36 words
The cell of claim 78 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the ribosome binding region and the translation initiation region of the RNA transcript produced by said gene.
Claim 88
Dependent Claim
15 words
The cell of claim 61 wherein said non-native polynucleotide construct is incorporated into a vector.
Claim 89
Dependent Claim
11 words
The cell of claim 85 wherein said vector is a plasmid.
Claim 90
Dependent Claim
12 words
The cell of claim 85 wherein said vector is a viral vector.
Claim 91
Dependent Claim
13 words
The cell of claim 85 wherein said vector is either single-stranded or double-stranded.
Claim 92
Dependent Claim
10 words
The cell of claim 61 wherein said cell is prokaryotic.
Claim 93
Dependent Claim
10 words
The cell of claim 61 wherein said cell is eukaryotic.
Claim 94
Dependent Claim
18 words
The cell of claim 61 wherein said non-native polynucleotide construct is introduced into the nucleus of said cell.
Claim 95
Dependent Claim
15 words
The cell of any one of claims 61 to 91 wherein said construct is DNA.
Claim 96
Dependent Claim
61 words
A cell comprising a non-native polynucleotide construct which construct produces in said cell a non-naturally occurring polynucleotide complementary to an RNA transcript produced by a gene in said cell, said non-naturally occurring polynucleotide further comprising a stable stem and loop structure with a negative ΔG of formation, whereby said non-naturally occurring polynculeotide regulates the function of said gene in said cell.
Claim 97
Dependent Claim
61 words
A cell comprising a non-native polynucleotide construct which construct produces in said cell a non-naturally occurring polynucleotide complementary to an RNA transcript produced by a gene in said cell, said non-naturally occurring polynucleotide further comprising a stable stem and loop structure with a negative ΔG of formation, whereby said non-naturally occurring polynculeotide regulates the function of said gene in said cell.
Claim 98
Dependent Claim
21 words
The cell of claim 93 wherein the stem and loop structure is at the 3' end of the non-naturally occurring polynucleotide.
Claim 99
Dependent Claim
21 words
The cell of claim 93 wherein the stem and loop structure is at the 5' end of the non-naturally occurring polynucleotide.
Claim 100
Dependent Claim
15 words
The cell of claim 93 wherein the ΔG of formation is at least -4.5 Kcal/mol.
Claim 101
Dependent Claim
15 words
The cell of claim 93 wherein the ΔG of formation is at least -12.5 Kcal/mol.
Claim 102
Dependent Claim
17 words
The cell of claim 93 wherein the stem and loop structure is derived from a prokaryotic RNA.
Claim 103
Dependent Claim
17 words
The cell of claim 93 wherein the stem and loop structure is derived from a eukaryotic RNA.
Claim 104
Dependent Claim
34 words
The cell of claim 93 wherein the stem and loop structure is derived from an RNA selected from the group consisting of tRNA, mRNA, 5S RNA, rRNA, hRNA, viroid RNA and viral genomic ssRNA.
Claim 105
Dependent Claim
30 words
The cell of claim 93 wherein the stem and loop structure polynucleotide is derived from an RNA selected from the group consisting of tRNA, mRNA, 5S RNA, rRNA and hnRNA.
Claim 106
Dependent Claim
16 words
The cell of claim 93 wherein the stem and loop structure is derived from a rRNA.
Claim 107
Dependent Claim
16 words
The cell of claim 93 wherein the stem and loop structure is derived from a tRNA.
Claim 108
Dependent Claim
16 words
The cell of claim 93 wherein the stem and loop structure is derived from a mRNA.
Claim 109
Dependent Claim
14 words
The cell of claim 93 wherein said non-native polynucleotide construct comprises an inducible promoter.
Claim 110
Dependent Claim
11 words
The cell of claim 93 wherein said gene is an oncogene.
Claim 111
Dependent Claim
12 words
The cell of claim 93 wherein said gene is a viral gene.
Claim 112
Dependent Claim
22 words
The cell of claim 93 wherein said gene encodes a positive regulator whose presence is necessary for the expression of another gene.
Claim 113
Dependent Claim
16 words
The cell of claim 93 wherein said gene is related to a genetic disease or defect.
Claim 114
Dependent Claim
11 words
The cell of claim 93 therein said gene encodes a protein.
Claim 115
Dependent Claim
28 words
The cell of claim 110 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the coding region of the RNA transcript produced by said gene.
Claim 116
Dependent Claim
28 words
The cell of claim 110 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the non-coding region of the RNA transcript produced by said gene.
Claim 117
Dependent Claim
29 words
The cell of claim 110 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the 5' non-coding region of the RNA transcript produced by said gene.
Claim 118
Dependent Claim
29 words
The cell of claim 110 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the ribosome binding region of the RNA transcript produced by said gene.
Claim 119
Dependent Claim
29 words
The cell of claim 110 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the translation initiation region of the RNA transcript produced by said gene.
Claim 120
Dependent Claim
34 words
The cell of claim 110 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the ribosome binding region and the translation initiation region of said RNA transcript produced by said gene.
Claim 121
Dependent Claim
15 words
The cell of claim 93 wherein the non-native polynucleotide construct is incorporated into a vector.
Claim 122
Dependent Claim
11 words
The cell of claim 117 wherein said vector is a plasmid.
Claim 123
Dependent Claim
12 words
The cell of claim 117 wherein said vector is a viral vector.
Claim 124
Dependent Claim
13 words
The cell of claim 117 wherein said vector is either single-stranded or double-stranded.
Claim 125
Dependent Claim
10 words
The cell of claim 93 wherein said cell is prokaryotic.
Claim 126
Dependent Claim
10 words
The cell of claim 93 wherein said cell is eukaryotic.
Claim 127
Dependent Claim
18 words
The cell of claim 93 wherein the non-native polynucleotide construct is introduced into the nucleus of said cell.
Claim 128
Dependent Claim
15 words
The cell of any one of claims 93 and 123 wherein said construct is DNA.
Claim 129
Independent Claim
106 words
A method of regulating the function of a gene in a cell, comprising the steps of: (a) preparing a non-native polynucleotide construct which comprises (i) a transcriptional promoter segment; (ii) a segment coding for a stable stem and loop structure with a negative ΔG of formation operably linked downstream of said promoter segment; and (iii) a polynucleotide segment comprising a gene segment operably linked downstream of said promoter segment and inverted with respect to a gene in a cell; and (b) introducing said non-native polynucleotide construct into said cell containing said gene; whereby the transcript of said inverted gene segment regulates the function of said gene.
Claim 130
Independent Claim
106 words
A method of regulating the function of a gene in a cell, comprising the steps of: (a) preparing a non-native polynucleotide construct which comprises (i) a transcriptional promoter segment; (ii) a segment coding for a stable stem and loop structure with a negative ΔG of formation operably linked downstream of said promoter segment; and (iii) a polynucleotide segment comprising a gene segment operably linked downstream of said promoter segment and inverted with respect to a gene in a cell; and (b) introducing said non-native polynucleotide construct into said cell containing said gene; whereby the transcript of said inverted gene segment regulates the function of said gene.
Claim 131
Dependent Claim
25 words
The method of claim 125 wherein the stem and loop structure is at the 3' end of the transcript produced from said inverted gene segment.
Claim 132
Dependent Claim
25 words
The method of claim 125 wherein the stem and loop structure is at the 5' end of the transcript produced from said inverted gene segment.
Claim 133
Dependent Claim
15 words
The method of claim 125 wherein the ΔG of formation is at least -4.5 Kcal/mol.
Claim 134
Dependent Claim
15 words
The method of claim 125 wherein the ΔG of formation is at least -12.5 Kcal/mol.
Claim 135
Dependent Claim
17 words
The method of claim 125 wherein the stem and loop structure is derived from a prokaryotic RNA.
Claim 136
Dependent Claim
17 words
The method of claim 125 wherein the stem and loop structure is derived from a eukaryotic RNA.
Claim 137
Dependent Claim
34 words
The method of claim 125 wherein the stem and loop structure is derived from an RNA selected from the group consisting of tRNA, mRNA, 5S RNA, rRNA, hnRNA, viroid RNA and viral genomic ssRNA.
Claim 138
Dependent Claim
29 words
The method of claim 125 wherein the stem and loop structure is derived from an RNA selected from the group consisting of tRNA, mRNA, 5S RNA, rRNA and hnRNA.
Claim 139
Dependent Claim
16 words
The method of claim 125 wherein the stem and loop structure is derived from a rRNA.
Claim 140
Dependent Claim
16 words
The method of claim 125 wherein the stem and loop structure is derived from a tRNA.
Claim 141
Dependent Claim
16 words
The method of claim 125 wherein the stem and loop structure is derived from a mRNA.
Claim 142
Dependent Claim
49 words
The method of regulating the function of a gene in a cell according to claim 125 wherein said transcriptional promoter segment of said non-native polynucleotide construct comprises an inducible promoter and which method further comprises the step of providing to said cell an inducer for inducing said inducible promoter.
Claim 143
Dependent Claim
22 words
The method of regulating the function of a gene in a cell according to claim 125 wherein said gene is an oncogene.
Claim 144
Dependent Claim
23 words
The method of regulating the function of a gene is a cell according to claim 125 wherein said gene is a viral gene.
Claim 145
Dependent Claim
33 words
The method of regulating the function of a gene in a cell according to claim 125 wherein said gene encodes a positive regulator whose presence is necessary for the expression of another gene.
Claim 146
Dependent Claim
27 words
The method of regulating the function of a gene in a cell according to claim 125 wherein said gene is related to a genetic disease or defect.
Claim 147
Dependent Claim
22 words
The method of regulating the function of a gene in a cell according to claim 125 wherein said gene encodes a protein.
Claim 148
Dependent Claim
41 words
The method of regulating the function of a gene in a cell according to claim 142 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to a coding region of the RNA transcript produced by said gene.
Claim 149
Dependent Claim
41 words
The method of regulating the function of a gene in a cell according to claim 142 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to a non-coding region of the RNA transcript produced by said gene.
Claim 150
Dependent Claim
42 words
The method of regulating the function of a gene in a cell according to claim 142 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the 5' non-coding region of the RNA transcript produced by said gene.
Claim 151
Dependent Claim
42 words
The method of regulating the function of a gene in a cell according to claim 142 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the ribosome binding region of the RNA transcript produced by said gene.
Claim 152
Dependent Claim
42 words
The method of regulating the function of a gene in a cell according to claim 142 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the translation initiation region of the RNA transcript produced by said gene.
Claim 153
Dependent Claim
47 words
The method of regulating the function of a gene in a cell according to claim 142 wherein said polynucleotide segment comprising a gene segment encodes an RNA transcript complementary to the ribosome binding region and the translation initiation region of the RNA transcript produced by said gene.
Claim 154
Dependent Claim
26 words
The method of regulating the function of a gene in a cell according to claim 125 wherein said non-native polynucleotide construct is incorporated into a vector.
Claim 155
Dependent Claim
22 words
The method of regulating the function of a gene in a cell according to claim 149 wherein said vector is a plasmid.
Claim 156
Dependent Claim
23 words
The method of regulating the function of a gene in a cell according to claim 149 wherein said vector is a viral vector.
Claim 157
Dependent Claim
24 words
The method of regulating the function of a gene in a cell according to claim 149 wherein said vector is either single-stranded or double-stranded.
Claim 158
Dependent Claim
10 words
The method of claim 125 wherein said cell is prokaryotic.
Claim 159
Dependent Claim
10 words
The method of claim 125 wherein said cell is eukaryotic.
Claim 160
Dependent Claim
18 words
The method of claim 125 wherein the non-native polynucleotide construct is introduced into the nucleus of said cell.
Claim 161
Dependent Claim
17 words
The method of claim 125 wherein the non-native polynucleotide construct is introduced into said cell by microinjection.
Claim 162
Dependent Claim
17 words
The method of claim 125 wherein the non-native polynucleotide construct is introduced into said cell by electroporation.
Claim 163
Dependent Claim
17 words
The method of claim 125 wherein the non-native polynucleotide construct is introduced into said cell by coprecipitation.
Claim 164
Independent Claim
71 words
A method of regulating the function of a gene in a cell, comprising introducing into a cell a non-native polynucleotide construct which, produces a non-naturally occurring polynucleotide complementary to an RNA transcript produced by said gene in said cell, said non-naturally occurring polynucleotide further comprising a stable stem and loop structure with a negative ΔG of formation, whereby said non-naturally occurring polynucleotide regulates the function of said gene, in said cell.
Claim 165
Independent Claim
71 words
A method of regulating the function of a gene in a cell, comprising introducing into a cell a non-native polynucleotide construct which, produces a non-naturally occurring polynucleotide complementary to an RNA transcript produced by said gene in said cell, said non-naturally occurring polynucleotide further comprising a stable stem and loop structure with a negative ΔG of formation, whereby said non-naturally occurring polynucleotide regulates the function of said gene, in said cell.
Claim 166
Dependent Claim
21 words
The method of claim 159 wherein the stem and loop structure is at the 3' end of the non-naturally occurring polynucleotide.
Claim 167
Dependent Claim
21 words
The method of claim 159 wherein the stem and loop structure is at the 5' end of the non-naturally occurring polynucleotide.
Claim 168
Dependent Claim
15 words
The method of claim 159 wherein the ΔG of formation is at least -4.5 Kcal/mol.
Claim 169
Dependent Claim
15 words
The method of claim 159 wherein the ΔG of formation of at least -12.5 Kcal/mol.
Claim 170
Dependent Claim
17 words
The method of claim 159 wherein the stem and loop structure is derived from a prokaryotic RNA.
Claim 171
Dependent Claim
17 words
The method of claim 159 wherein the stem and loop structure is derived from a eukaryotic RNA.
Claim 172
Dependent Claim
34 words
The method of claim 159 wherein the stem and loop structure is derived from an RNA selected from the group consisting of tRNA, mRNA, 5S RNA, rRNA, hRNA, viroid RNA and viral genomic ssRNA.
Claim 173
Dependent Claim
29 words
The method of claim 159 wherein the stem and loop structure is derived from an RNA selected from the group consisting of tRNA, mRNA, 5S RNA, rRNA and hnRNA.
Claim 174
Dependent Claim
16 words
The method of claim 159 wherein the stem and loop structure is derived from a rRNA.
Claim 175
Dependent Claim
16 words
The method of claim 159 wherein the stem and loop structure is derived from a tRNA.
Claim 176
Dependent Claim
16 words
The method of claim 159 wherein the stem and loop structure is derived from a mRNA.
Claim 177
Dependent Claim
44 words
The method of regulating the function of a gene in a cell according to claim 159 wherein said non-native polynucleotide construct comprises an inducible promoter and which method further comprises the step of providing to said cell an inducer for inducing said inducible promoter.
Claim 178
Dependent Claim
22 words
The method of regulating the function of a gene in a cell according to claim 159 wherein said gene is an oncogene.
Claim 179
Dependent Claim
23 words
The method of regulating the function of a gene in a cell according to claim 159 wherein said gene is a viral gene.
Claim 180
Dependent Claim
33 words
The method of regulating the function of a gene in a cell according to claim 159 wherein said gene encodes a positive regulator whose presence is necessary for the expression of another gene.
Claim 181
Dependent Claim
27 words
The method of regulating the function of a gene in a cell according to claim 159 wherein said gene is related to a genetic disease or defect.
Claim 182
Dependent Claim
22 words
The method of regulating the function of a gene in a cell according to claim 159 wherein said gene encodes a protein.
Claim 183
Dependent Claim
39 words
The method of regulating the function of a gene in a cell according to claim 176 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the coding region of the RNA transcript produced by said gene.
Claim 184
Dependent Claim
39 words
The method of regulating the function of a gene in a cell according to claim 176 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the non-coding region of the RNA transcript produced by said gene.
Claim 185
Dependent Claim
40 words
The method of regulating the function of a gene in a cell according to claim 176 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the 5' non-coding region of the RNA transcript produced by said gene.
Claim 186
Dependent Claim
40 words
The method of regulating the function of a gene in a cell according to claim 176 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the ribosome binding region of the RNA transcript produced by said gene.
Claim 187
Dependent Claim
40 words
The method of regulating the function of a gene is a cell according to claim 176 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the translation initiation region of the RNA transcript produced by said gene.
Claim 188
Dependent Claim
45 words
The method of regulating the function of a gene in a cell according to claim 176 wherein the polynucleotide produced from the non-native polynucleotide construct is complementary to the ribosome binding region and the translation initiation region of the RNA transcript produced by said gene.
Claim 189
Dependent Claim
26 words
The method of regulating the function of a gene in a cell according to claim 159 wherein said non-native polynucleotide construct is incorporated into a vector.
Claim 190
Dependent Claim
22 words
The method of regulating the function of a gene is a cell according to claim 183 wherein said vector is a plasmid.
Claim 191
Dependent Claim
23 words
The method of regulating the function of a gene in a cell according to claim 183 wherein said vector is a viral vector.
Claim 192
Dependent Claim
24 words
The method of regulating the function of a gene in a cell according to claim 183 wherein said vector is either single-stranded or double-stranded.
Claim 193
Dependent Claim
10 words
The method of claim 159 wherein said cell is prokaryotic.
Claim 194
Dependent Claim
10 words
The method of claim 159 wherein said cell is eukaryotic.
Claim 195
Dependent Claim
18 words
The method of claim 159 wherein the non-native polynucleotide construct is introduced into the nucleus of said cell.
Claim 196
Dependent Claim
17 words
The method of claim 159 wherein the non-native polynucleotide construct is introduced into said cell by microinjection.
Claim 197
Dependent Claim
17 words
The method of claim 159 wherein the non-native polynucleotide construct is introduced into said cell by electroporation.
Claim 198
Dependent Claim
17 words
The method of claim 159 wherein the non-native polynucleotide construct is introduced into said cell by coprecipitation.
Claim 199
Independent Claim
17 words
The method of any one of claims 125 to 158 wherein said non-native polynucleotide construct is DNA.
Claim 200
Independent Claim
17 words
The method of any one of claims 159 to 192 wherein said non-native polynucleotide construct is RNA.
Claim 201
Independent Claim
17 words
The method of any one of claims 159 to 192 wherein said non-native polynucleotide construct is DNA.
Claim 202
Independent Claim
54 words
A method of regulating the function of a gene in a cell which comprises: introducing into said cell the polynucleotide construct of any one of claims 1 to 28 whereby the transformed cell is obtained; and growing said transformed cell whereby the transcript of the inverted gene segment regulates the function of said gene.
Claim 203
Independent Claim
51 words
A method of regulating the function of a gene in a cell which comprises: introducing into said cell the polynucleotide construct of any one of claims 31 to 58 whereby the transformed cell is obtained; and growing said transformed cell whereby said non-naturally occurring polynucleotide regulates the function of said gene.
Claim 204
Dependent Claim
23 words
The cell of claim 61 or 93 wherein said non-native polynucleotide is incorporated in or associated with the native DNA of said cell.
References Cited (Backward Citations)
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Cited
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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) | 1996-05-03 | 1996-11-02 | 1997-05-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-05-03 | 2000-11-02 | 2001-05-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-05-03 | 2004-11-02 | 2005-05-04 | $7700.00 | Window Closed / Lapsed |