US4072574A
Expired / Lapsed
US
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System for in vitro assay for mutagenicity and/or carcinogenicity of chemicals or other exogenous agents
Assignee: Institute for Cancer Research
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Filing: 1976-07-30
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Granted: 1978-02-07
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Expires: 1996-07-30
Abstract
Abstract This involves an in vitro assay system to screen for mutagenic and/or carcinogenic agents based on alterations in the fidelity of DNA synthesis. In the system, chemicals will be tested for their mutagenicity or carcinogenicity by measuring increases in the number of mistakes incorporated by purified DNA polymerases using synthetic polynucleotide templates. This system offers the advantage of performing this analysis in the test tube so that all parameters in the reaction can be monitored. Agents that enhance misincorporation during DNA synthesis would be predicted to have a higher probability of being mutagens or carcinogens in the human population.
Inventors
- 👤 Lawrence A. Loeb
- 👤 Michael A. Sirover
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 19:14
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/48
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
76 words
The method of testing in vitro for the likelihood of mutagenic or carcinogenic properties in a given substance which comprises copying polynucleotides of known composition in the presence of the substance by means of DNA polymerases in a solution which also contains correct complementary nucleotides for said polynucleotides and non-complementary nucleotides for said polynucleotides, and also doing the same incorrect copying in the absence of the substance, and measuring the relative accuracy of the respective copying.
Claim 2
Independent Claim
76 words
The method of testing in vitro for the likelihood of mutagenic or carcinogenic properties in a given substance which comprises copying polynucleotides of known composition in the presence of the substance by means of DNA polymerases in a solution which also contains correct complementary nucleotides for said polynucleotides and non-complementary nucleotides for said polynucleotides, and also doing the same incorrect copying in the absence of the substance, and measuring the relative accuracy of the respective copying.
Claim 3
Independent Claim
94 words
The method of testing a substance for the likelihood of mutagenic or carcinogenic properties which comprises replicating a polynucleotide by having a DNA polymerase copy a polynucleotide of known composition in the absence of the substance in a solution including both a correct complementary and an incorrect non-complementary nucleotide for said polynucleotide, also replicating the same polynucleotide by having the same polymerase copy said polynucleotide in the presence of that substance in a solution including the same correct and incorrect nucleotides for said polynucleotides and comparing the results from the standpoint of producing mutations.
Claim 4
Independent Claim
94 words
The method of testing a substance for the likelihood of mutagenic or carcinogenic properties which comprises replicating a polynucleotide by having a DNA polymerase copy a polynucleotide of known composition in the absence of the substance in a solution including both a correct complementary and an incorrect non-complementary nucleotide for said polynucleotide, also replicating the same polynucleotide by having the same polymerase copy said polynucleotide in the presence of that substance in a solution including the same correct and incorrect nucleotides for said polynucleotides and comparing the results from the standpoint of producing mutations.
Claim 5
Independent Claim
99 words
The method of testing for the likelihood of mutagenic or carcinogenic properties which comprises including in a solution of a polynucleotide, a nucleotide which is a correct nucleotide for that particular polynucleotide, a nucleotide which is not a correct nucleotide for that particular polynucleotide and a polymerase; replicating the polynucleotide in said solution in which a substance to be tested for its likelihood of mutagenic or carcinogenic properties is also included also replicating the polynucleotide in an identical solution in which said substance to be tested is not included and comparing the results from the standpoint of producing mutations.
Claim 6
Independent Claim
99 words
The method of testing for the likelihood of mutagenic or carcinogenic properties which comprises including in a solution of a polynucleotide, a nucleotide which is a correct nucleotide for that particular polynucleotide, a nucleotide which is not a correct nucleotide for that particular polynucleotide and a polymerase; replicating the polynucleotide in said solution in which a substance to be tested for its likelihood of mutagenic or carcinogenic properties is also included also replicating the polynucleotide in an identical solution in which said substance to be tested is not included and comparing the results from the standpoint of producing mutations.
Claim 7
Dependent Claim
15 words
The method of claim 3, in which Mg 2+ is also included in the solution.
Claim 8
Dependent Claim
33 words
The method of claim 3, in which there is used a synthetic polynucleotide of defined composition which may be copied with procaryotic, eucaryotic or viral DNA polymerases, and such a polymerase is used.
Claim 9
Dependent Claim
15 words
The method of claim 5, in which Mg 2+ is also included in the solution.
Claim 10
Dependent Claim
34 words
A method of claim 5, in which the correct nucleotide is labeled with [α- 32 P] of low specific activity and the incorrect nucleotide is labeled with [ 3 H] of high specific activity.
Claim 11
Independent Claim
71 words
The method of testing in vitro a substance for the likelihood of mutagenic and carcinogenic properties, which comprises relicating a polynucleotide with a DNA polymerase in a solution containing a correct complementary and an incorrect non-complementary nucleotide for said polynucleotide in the absence of said substance, also replicating that polynucleotide by that same polymerase in a solution containing that substance, and comparing the results from the standpoint of production of mutations.
Claim 12
Independent Claim
71 words
The method of testing in vitro a substance for the likelihood of mutagenic and carcinogenic properties, which comprises relicating a polynucleotide with a DNA polymerase in a solution containing a correct complementary and an incorrect non-complementary nucleotide for said polynucleotide in the absence of said substance, also replicating that polynucleotide by that same polymerase in a solution containing that substance, and comparing the results from the standpoint of production of mutations.
Claim 13
Independent Claim
142 words
The method of testing a particular substance for likelihood of mutagenicity or carcinogenicity which comprises (a) synthesizing in vitro a particular polynucleotide which contains a particular first nucleotide and does not contain a particular second nucleotide, in a solution which contains as a template the same polynucleotide as is being synthesized and includes also the first nucleotide, the second nucleotide, a particular polymerase and the particular substance to be tested; also (b) synthesizing in vitro the same particular polynucleotide in a solution which contains as a template the same polynucleotide and includes also the first nucleotide, the second nucleotide and the same polymerase but does not include the particular substance to be tested; and (c) determining whether or not the proportion of second nucleotide to first nucleotide in the product of synthesis (a) significantly exceeds that in the product of synthesis (b).
Claim 14
Independent Claim
142 words
The method of testing a particular substance for likelihood of mutagenicity or carcinogenicity which comprises (a) synthesizing in vitro a particular polynucleotide which contains a particular first nucleotide and does not contain a particular second nucleotide, in a solution which contains as a template the same polynucleotide as is being synthesized and includes also the first nucleotide, the second nucleotide, a particular polymerase and the particular substance to be tested; also (b) synthesizing in vitro the same particular polynucleotide in a solution which contains as a template the same polynucleotide and includes also the first nucleotide, the second nucleotide and the same polymerase but does not include the particular substance to be tested; and (c) determining whether or not the proportion of second nucleotide to first nucleotide in the product of synthesis (a) significantly exceeds that in the product of synthesis (b).
References Cited (Backward Citations)
US3816263A
Cited
US3850749A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1981-02-06 | 1981-08-08 | 1982-02-07 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-02-06 | 1985-08-08 | 1986-02-07 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-02-06 | 1989-08-08 | 1990-02-07 | $7700.00 | Window Closed / Lapsed |