US5180818A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Site specific cleavage of single-stranded dna
Assignee: University Research Corp
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Filing: 1990-03-21
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Granted: 1993-01-19
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Expires: 2010-03-21
Abstract
Abstract Method for specifically cleaving a single stranded DNA molecule. The method includes providing an RNA molecule having a deoxyribonuclease activity independent of any protein, and contacting that RNA molecule with the single stranded DNA molecule to cause the single stranded DNA molecule to be cleaved.
Inventors
- 👤 Thomas R. Cech
- 👤 Daniel Herschlag
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 22:54
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/113; C12N2310/00; C12N2310/10; C12N2310/12; C12N2310/124; C12N2310/1241; C12N2310/126
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
111 words
A method for specifically cleaving a single-stranded DNA molecule, comprising the steps of: (a) providing a first RNA molecule selected from the group consisting of a group I intron, a group II intron, or RNAse P, that cleaves a second RNA molecule to leave a 3'-OH, said first RNA molecule having a deoxyribonuclease activity, and (b) contacting said first RNA molecule with said single-stranded DNA molecule under conditions which allow said first RNA molecule to cause said single-stranded DNA molecule to be cleaved said conditions including providing Mg 2+ ions and guanosine or guanosine triphosphate at a pH between 6.0 and 9.5 and a temperature between 20° C. and 70° C.
Claim 2
Independent Claim
111 words
A method for specifically cleaving a single-stranded DNA molecule, comprising the steps of: (a) providing a first RNA molecule selected from the group consisting of a group I intron, a group II intron, or RNAse P, that cleaves a second RNA molecule to leave a 3'-OH, said first RNA molecule having a deoxyribonuclease activity, and (b) contacting said first RNA molecule with said single-stranded DNA molecule under conditions which allow said first RNA molecule to cause said single-stranded DNA molecule to be cleaved said conditions including providing Mg 2+ ions and guanosine or guanosine triphosphate at a pH between 6.0 and 9.5 and a temperature between 20° C. and 70° C.
Claim 3
Dependent Claim
36 words
The method of claim 1, further comprising providing said RNA molecule in a reaction medium at a concentration sufficient to cause cleavage of at least 1% of a population of the DNA molecules in an hour.
Claim 4
Dependent Claim
36 words
The method of claim 1, further comprising providing said RNA molecule in a reaction medium at a concentration sufficient to cause cleavage of at least 10% of a population of the DNA molecules in an hour.
Claim 5
Dependent Claim
22 words
The method of claim 1, wherein said RNA molecule comprises the portions of an RNA molecule of Tetrahymena having said deoxyribonuclease activity.
Claim 6
Dependent Claim
25 words
The method of claim 4, wherein said RNA molecule is L-19, L-21, or an RNA molecule comprising the portions of L-19 having said deoxyribonuclease activity.
Claim 7
Dependent Claim
30 words
The method of claim 1, further comprising the step of providing a chelator of said Mg 2+ ions to stop further reaction of said RNA molecule with said DNA molecule.
Claim 8
Dependent Claim
12 words
The method of claim 6, wherein said chelator is ethylenediaminetetraacetic acid (EDTA).
Claim 9
Dependent Claim
33 words
The method of claim 1, wherein said RNA molecule comprises a binding site for single-stranded DNA, which binding site is complementary to nucleotides adjacent to a cleavage site on said single-stranded DNA molecule.
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) | 1996-01-19 | 1996-07-20 | 1997-01-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-01-19 | 2000-07-20 | 2001-01-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-01-19 | 2004-07-20 | 2005-01-19 | $7700.00 | Window Closed / Lapsed |