US4786600A
Expired / Lapsed
US
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Autocatalytic replication of recombinant RNA
Assignee: Columbia University
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Filing: 1984-05-25
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Granted: 1988-11-22
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Expires: 2004-05-25
Abstract
Abstract This invention concerns recombinant RNA molecules comprising a recognition sequence for the binding of an RNA-directed RNA polymerase, a sequence for the initiation of product strand synthesis and a heterologous sequence of interest inserted at a specific site in the internal region of the recombinant molecule. Such recombinant RNA molecules are capable of serving as a template for the synthesis of complementary single-stranded molecules by RNA-directed RNA polymerase. The product molecules so formed are also capable of serving as a template for the synthesis of additional copies of the original recombinant RNA molecule. In a preferred embodiment of the invention Qβ replicase is used as the RNA-directed RNA polymerase.
Inventors
- 👤 Fred R. Kramer
- 👤 Eleanor A. Miele
- 👤 Donald R. Mills
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 10:11
CPC Classifications
C; C12; C12N; C12N7/00; C12N15/00; C12N15/09; C12N15/10; C12P; C12P19/00; C12P19/26; C12P19/28; C12P19/30; C12P19/34; C12Q; C12Q1/00; C12Q1/68; C12Q1/6844; C12Q1/6867; C12N2795/00; C12N2795/00011; C12N2795/10011; C12N2795/10111; C12N2795/10122; C12N2795/18011; C12N2795/18111; C12N2795/18122
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
22 words
A recombinant single-stranded RNA molecule, wherein the sequence required for the initiation of product strand synthesis is a cytidine-rich 3' terminal sequence.
Claim 2
Dependent Claim
22 words
A recombinant single-stranded RNA molecule, wherein the sequence required for the initiation of product strand synthesis is a cytidine-rich 3' terminal sequence.
Claim 3
Dependent Claim
16 words
A recombinant single-stranded RNA molecule as in claim 1 which comprises at least one radiolabeled nucleotide.
Claim 4
Dependent Claim
30 words
A recombinant single-stranded RNA molecule as in claim 1, wherein the insertion site for the heterologous sequence of interest is in a loop where viable mutations are known to occur.
Claim 5
Dependent Claim
28 words
A recombinant single-stranded RNA molecule as in claim 3, wherein the insertion site is at or near a guanosine residue hypersusceptible to cleavage by ribonuclease T 1 .
Claim 6
Dependent Claim
17 words
A recombinant single-stranded RNA molecule as in claim 1, wherein the MDV-1 RNA is MDV-1 (+) RNA.
Claim 7
Dependent Claim
17 words
A recombinant single-stranded RNA molecule as in claim 1, wherein the MDV-1 RNA is MDV-1 (-) RNA.
Claim 8
Dependent Claim
22 words
A recombinant single-stranded RNA molecule as in claim 5, wherein the heterologous sequence of interest is inserted between nucleotides 63 and 64.
Claim 9
Dependent Claim
19 words
A recombinant single-stranded RNA molecule as in claim 7, wherein the inserted heterologous sequence of interest is decaadenylic acid.
Claim 10
Dependent Claim
116 words
A recombinant, single-stranded RNA molecule capable of serving as a template in vitro for the synthesis of a complementary single-stranded RNA molecule by Qβ replicase which comprises MDV-1 RNA having the structure shown in FIG. 2 or a mutant thereof and a heterologous RNA sequence inserted into the MDV-1 RNA or the mutant thereof essentially without changing the secondary or tertiary structure of the MDV-1 RNA, the heterologous sequence of interest being inserted between two nucleotides, neither of the nucleotides being (a) hydrogen-bonded to any other nucleotide within the MDV-1 RNA, or (b) present within a Qβ replicase recognition sequence, or (c) present within a sequence for the initiation of product strand synthesis by Qβ replicase.
Claim 11
Dependent Claim
116 words
A recombinant, single-stranded RNA molecule capable of serving as a template in vitro for the synthesis of a complementary single-stranded RNA molecule by Qβ replicase which comprises MDV-1 RNA having the structure shown in FIG. 2 or a mutant thereof and a heterologous RNA sequence inserted into the MDV-1 RNA or the mutant thereof essentially without changing the secondary or tertiary structure of the MDV-1 RNA, the heterologous sequence of interest being inserted between two nucleotides, neither of the nucleotides being (a) hydrogen-bonded to any other nucleotide within the MDV-1 RNA, or (b) present within a Qβ replicase recognition sequence, or (c) present within a sequence for the initiation of product strand synthesis by Qβ replicase.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-11-22 | 1992-05-23 | 1992-11-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-11-22 | 1996-05-23 | 1996-11-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-11-22 | 2000-05-23 | 2000-11-22 | $7700.00 | Window Closed / Lapsed |