Claims
- 1. A targeting construct comprising:
(a) a first polynucleotide sequence homologous to at least a first portion of a GPRC5B-like gene, wherein the GPRC5B-like gene comprises SEQ ID NO:1; (b) a second polynucleotide sequence homologous to at least a second portion of the GPRC5B-like gene; and (c) a selectable marker.
- 2. A method of producing a targeting construct, the method comprising:
(a) providing a first polynucleotide sequence homologous to at least a first portion of a GPRC5B-like gene, wherein the GPRC5B-like gene comprises SEQ ID NO:1; (b) providing a second polynucleotide sequence homologous to at least a second portion of the GPRC5B-like gene; (c) providing a selectable marker; and (d) inserting the first sequence, second sequence, and selectable marker into a vector, to produce the targeting construct.
- 3. A cell comprising a disruption in a GPRC5B-like gene, wherein the GPRC5B-like gene comprises SEQ ID NO:1.
- 4. The cell of claim 3, wherein the cell is a murine cell.
- 5. The cell of claim 4, wherein the murine cell is an embryonic stem cell.
- 6. A non-human transgenic animal comprising a disruption in a GPRC5B-like gene, wherein the GPRC5B-like gene comprises SEQ ID NO:1.
- 7. The non-human transgenic animal of claim 6, wherein the transgenic animal is a mouse.
- 8. A cell derived from the transgenic mouse of claim 7.
- 9. A method of producing a transgenic mouse comprising a disruption in a GPRC5B-like gene, the method comprising:
(a) introducing the targeting construct of claim 1 into a cell; (b) introducing the cell into a blastocyst; (c) implanting the resulting blastocyst into a pseudopregnant mouse, wherein said pseudopregnant mouse gives birth to a chimeric mouse; and (d) breeding the chimeric mouse to produce the transgenic mouse.
- 10. A method of identifying an agent that modulates the expression or function of a GPRC5B-like gene, the method comprising:
(a) providing a non-human transgenic animal comprising a disruption in a GPRC5B-like gene, wherein the GPRC5B-like gene comprises SEQ ID NO:1; (b) administering an agent to the non-human transgenic animal; and (c) determining whether the expression or function of the disrupted GPRC5B-like gene in the non-human transgenic animal is modulated.
- 11. A method of identifying an agent that modulates the expression or function of a GPRC5B-like gene, the method comprising:
(a) providing a cell comprising a disruption in a GPRC5B-like gene; (b) contacting the cell with the agent; and (c) determining whether the expression or function of the GPRC5B-like gene is modulated.
- 12. The method of claim 11, wherein the cell is derived from the non-human transgenic animal of claim 6.
- 13. An agent identified by the method of claim 10 or claim 11.
- 14. A transgenic mouse comprising a disruption in a GPRC5B-like gene, wherein there is no significant expression of the GPRC5B-like gene in the transgenic mouse.
- 15. A transgenic mouse comprising a homozygous disruption in a GPRC5B-like gene, wherein the transgenic mouse exhibits abnormal pain threshold.
- 16. The transgenic mouse of claim 15, wherein the transgenic mouse exhibits an increased pain threshold.
- 17. The transgenic mouse of claim 16, wherein the increased pain threshold is characterized by an increased latency to lick a hindpaw in response to a hot plate in a hot plate test, relative to a wild-type mouse.
- 18. A cell derived from the transgenic mouse of claim 14.
- 19. A method of identifying an agent that ameliorates a phenotype associated with a disruption in a GPRC5B-like gene, the method comprising:
(a) administering an agent to a transgenic mouse comprising a disruption in a GPRC5B-like gene, wherein the GPRC5B-like gene comprises SEQ ID NO:1; and (b) determining whether the agent has an affect on pain threshold.
- 20. An agent identified by the method of claim 19
- 21. A method of identifying an agent that has an affect on pain sensitivity, the method comprising:
(a) providing a mouse expressing a GPRC5B-like gene, wherein the GPRC5B-like gene comprises SEQ ID NO:1; (b) contacting the cell with a putative agent; and (c) determining whether the agent has an affect on pain sensitivity in the mouse.
- 22. A method of identifying an agent that inhibits the activity, expression, or function of a GPRC5B-like gene, the method comprising:
(a) providing a cell expressing a GPRC5B-like gene; (b) contacting the cell with an putative agent; and (c) determining whether the putative agent has an affect on activity, expression, or function of the GPRC5B-like gene, wherein the agent has an affect on pain threshold.
- 23. An agent identified by the method of claim 21 or claim 22.
- 24. A method of treating pain, the method comprising administering to a subject in need a therapeutically effective amount of an agent that inhibits the activity or function of a GPRC5B-like protein, wherein the GPRC5B-like protein comprises SEQ ID NO:2.
- 25. A method of screening for biologically active agents, the method comprising:
(a) combining a putative agent with a mammalian GPRC5B-like polypeptide; and (b) detecting an effect of the agent on GPRC5B-like polypeptide activity; wherein detection of a decrease or an increase in GPRC5B-like polypeptide activity is indicative of a biologically active agent.
- 26. A method of screening for biologically active agents, the method comprising:
(a) combining a putative agent with an isolated cell comprising a nucleic acid encoding a mammalian GPRC5B-like gene or a GPRC5B-like promoter sequence operably linked to a reporter gene; and (b) detecting an effect of the agent on GPRC5B-like activity; wherein detection of a decrease or an increase in GPRC5B activity is indicative of a biologically active agent.
- 27. A method of screening for biologically active agents, the method comprising:
(a) combining a putative agent with a non-human transgenic model comprising an exogenous and stably transfected mammalian GPRC5B-like gene or GPRC5B-like promoter sequence operably linked to a reporter gene; and (b) detecting an effect of the agent on GPRC5B-like function; wherein detection of a decrease or an increase in GPRC5B function is indicative of a biologically active agent.
- 28. The method of claim 27, wherein the agent has an effect on pain threshold in the non-human transgenic model.
- 29. An agonist or antagonist of a GPRC5B-like protein encoded by SEQ ID NO:1.
- 30. Phenotypic data associated with a transgenic mouse comprising a disruption in a GPRC5B-like gene, wherein the phenotypic data is in an electronic database.
RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional Application No. 60/256,199, filed Dec. 13, 2000, and U.S. Provisional Application No. 60/280,359, filed Mar. 29, 2001, the entire contents of each are incorporated herein by reference.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60256199 |
Dec 2000 |
US |
|
60280359 |
Mar 2001 |
US |