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Last Updated: September 19, 2020

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Claims for Patent: 5,180,818

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Summary for Patent: 5,180,818
Title: Site specific cleavage of single-stranded DNA
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.
Inventor(s): Cech; Thomas R. (Boulder, CO), Herschlag; Daniel (Boulder, CO)
Assignee: The University of Colorado Foundation, Inc. (Boulder, CO)
Application Number:07/496,852
Patent Claims:1. 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.sup.2+ ions and guanosine or guanosine triphosphate at a pH between 6.0 and 9.5 and a temperature between 20.degree. C. and 70.degree. C.

2. 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.

3. 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.

4. The method of claim 1, wherein said RNA molecule comprises the portions of an RNA molecule of Tetrahymena having said deoxyribonuclease activity.

5. 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.

6. The method of claim 1, further comprising the step of providing a chelator of said Mg.sup.2+ ions to stop further reaction of said RNA molecule with said DNA molecule.

7. The method of claim 6, wherein said chelator is ethylenediaminetetraacetic acid (EDTA).

8. 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.

Details for Patent 5,180,818

Applicant Tradename Biologic Ingredient Dosage Form BLA Number Approval Date Patent No. Assignee Estimated Patent Expiration Status Orphan Source
Schering INTRON A interferon alfa-2b VIAL 103132 001 1986-06-04   Start Trial The University of Colorado Foundation, Inc. (Boulder, CO) 2039-02-26 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 002 1986-06-04   Start Trial The University of Colorado Foundation, Inc. (Boulder, CO) 2039-02-26 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 003 1986-06-04   Start Trial The University of Colorado Foundation, Inc. (Boulder, CO) 2039-02-26 RX search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

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