US5071773A
Expired / Lapsed
US
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Hormone receptor-related bioassays
Assignee: Salk Institute for Biological Studies
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Filing: 1987-10-20
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Granted: 1991-12-10
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Expires: 2007-10-20
Abstract
Abstract The present invention discloses two hormone receptor-related bioassays. The first bioassay is useful for determining whether a protein suspected of being a hormone receptor has transcription-activating properties of a hormone receptor. The second bioassay is useful for evaluating whether compounds are functional ligands for receptor proteins. According to the first bioassay, cells that contain non-endogenous DNA which expresses a protein suspected of being a hormone receptor and which contain a DNA sequence encoding an operative hormone responsive promoter/enhancer element linked to an operative reporter gene, are cultured, the culturing being conducted in a culture medium containing a known hormone, or an analog thereof. The cultured cells are then monitored for induction of the product of the reporter gene as an indication of functional transcription-activating binding between the hormone or hormone analog and the protein suspected of being a hormone receptor. According to the second bioassay, cells that contain non-endogenous DNA which expresses hormone receptor or a functional engineered or modified form thereof, and which also contain a DNA sequence encoding an operative hormone responsive promoter/enhancer element linked to an operative reporter gene, are cultured, the culturing being conducted in culture medium containing at least one compound whose ability to functionally bind the receptor protein is sought to be determined. The cultured cells are then monitored for induction of the product of the report gene as an indicator of functional binding between the compound and the receptor.
Inventors
- 👤 Ronald M. Evans
- 👤 Cary A. Weinberger
- 👤 Stanley M. Hollenberg
- 👤 Vincent Giguere
- 👤 Jeffrey Arriza
- 👤 Catherine C. Thompson
- 👤 Estelita S. Ong
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 01:05
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/67; C07; C07K; C07K14/00; C07K14/435; C07K14/705; C07K14/72; C07K14/721; C07K14/723; C12Q; C12Q1/00; C12Q1/68; C12Q1/6897
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
124 words
A bioassay for determining whether (1) a protein (R) suspected of being a hormone receptor, or (2) a functional engineered or modified form (r) thereof, has transcription-activating properties of a hormone receptor, said bioassay comprising: (a) culturing cells containing non-endogenous DNA which expresses (R) or (r) and which contains a DNA sequence encoding an operative hormone response element linked to an operative reporter gene, the culturing being conducted in a culture medium containing a known hormone (H), or an analog thereof (aH) known to have transcription activating properties of said hormone, (H), and (b) monitoring in said cells induction of the product of said reporter gene as an indication of functional transcription-activating binding between hormone (H) or hormone analog (aH) with (R) or (r).
Claim 2
Dependent Claim
124 words
A bioassay for determining whether (1) a protein (R) suspected of being a hormone receptor, or (2) a functional engineered or modified form (r) thereof, has transcription-activating properties of a hormone receptor, said bioassay comprising: (a) culturing cells containing non-endogenous DNA which expresses (R) or (r) and which contains a DNA sequence encoding an operative hormone response element linked to an operative reporter gene, the culturing being conducted in a culture medium containing a known hormone (H), or an analog thereof (aH) known to have transcription activating properties of said hormone, (H), and (b) monitoring in said cells induction of the product of said reporter gene as an indication of functional transcription-activating binding between hormone (H) or hormone analog (aH) with (R) or (r).
Claim 3
Dependent Claim
145 words
A bioassay for evaluating whether compounds are functional ligands for receptor protein (R), or a functional engineered or modified form (r) thereof, wherein said functional engineered or modified form (r) has transcription-activating properties of receptor protein (R), said bioassay comprising: (a) culturing cells which contain non-endogenous DNA which expresses (R) or a functional engineered or modified form (r) thereof, wherein said functional engineered or modified form (r) has transcription-activating properties of (R), and which cells also contain a DNA sequence encoding an operative hormone response element linked to an operative reporter gene, the culturing being conducted in culture medium containing at least one compound (C) whose ability to functionally bind (R) or (r) is sought to be determined, and (b) monitoring in said cells induction of the product of said report gene as an indicator or functional binding between compound (C) and (R) or (r).
Claim 4
Dependent Claim
145 words
A bioassay for evaluating whether compounds are functional ligands for receptor protein (R), or a functional engineered or modified form (r) thereof, wherein said functional engineered or modified form (r) has transcription-activating properties of receptor protein (R), said bioassay comprising: (a) culturing cells which contain non-endogenous DNA which expresses (R) or a functional engineered or modified form (r) thereof, wherein said functional engineered or modified form (r) has transcription-activating properties of (R), and which cells also contain a DNA sequence encoding an operative hormone response element linked to an operative reporter gene, the culturing being conducted in culture medium containing at least one compound (C) whose ability to functionally bind (R) or (r) is sought to be determined, and (b) monitoring in said cells induction of the product of said report gene as an indicator or functional binding between compound (C) and (R) or (r).
Claim 5
Dependent Claim
117 words
A bioassay for determining whether a protein has transcription-activating properties of a hormone receptor, said bioassay comprising: culturing host cells containing reporter plasmid comprising an operative hormone response element functionally linked to an operative reporter gene, and expression plasmid comprising a DNA sequence encoding protein (R) or an engineered or modified form (r) thereof wherein (r) has transcription-activating properties of a receptor hormone, said culturing being done in the presence and absence of a hormone or analog thereof, said hormone or analog being able to activate said hormone response element in the presence of a protein that functions as a receptor for said hormone or analog thereof; and monitoring in said cells induction of reporter gene product.
Claim 6
Dependent Claim
117 words
A bioassay for determining whether a protein has transcription-activating properties of a hormone receptor, said bioassay comprising: culturing host cells containing reporter plasmid comprising an operative hormone response element functionally linked to an operative reporter gene, and expression plasmid comprising a DNA sequence encoding protein (R) or an engineered or modified form (r) thereof wherein (r) has transcription-activating properties of a receptor hormone, said culturing being done in the presence and absence of a hormone or analog thereof, said hormone or analog being able to activate said hormone response element in the presence of a protein that functions as a receptor for said hormone or analog thereof; and monitoring in said cells induction of reporter gene product.
Claim 7
Dependent Claim
26 words
A bioassay according to any of claims 1, 2 or 3 wherein said host cells are selected from the group consisting of CV-1 and COS cells.
Claim 8
Dependent Claim
20 words
A bioassay according to claim 3 wherein said reporter and expression plasmids also contain the origin of replication of SV-40.
Claim 9
Dependent Claim
17 words
A bioassay according to claim 3 wherein said reporter and expression plasmids also contain a selectable marker.
Claim 10
Dependent Claim
42 words
A bioassay according to any of claims 1, 2 or 3 wherein said hormone response element is selected from the group consisting of mouse tumor virus long terminal repeat (MTV LTR) and mammalian growth hormone promoter, and said reporter gene is CAT.
Claim 11
Dependent Claim
32 words
A bioassay according to any of claims 1, 2 and 3 wherein said protein (R) or said modified functional form (r) thereof is selected from the group consisting of steroid hormone receptors.
References Cited (Backward Citations)
US4859609A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1994-12-09 | 1995-06-10 | 1995-12-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-12-09 | 1999-06-10 | 1999-12-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-12-09 | 2003-06-10 | 2003-12-10 | $7700.00 | Window Closed / Lapsed |