Claims
- 1. A method for identifying a compound that modulates the level of a lipid or lipoprotein in a nematode comprising:
(a) contacting a compound with test nematodes; (b) comparing a phenotype of said test nematodes with said phenotype of nematodes not contacted with said compound, wherein a modification of said phenotype correlates with a modulated level of a lipid or lipoprotein, said phenotype being selected from the group consisting of (i) length of defecation cycle; (ii) rate of germ line development relative to rate of soma development; (iii) rate of embryonic development; and (iv) rate of post-embryonic development, whereby a difference in said phenotypes identifies said compound.
- 2. A method for isolating a gene that modulates the level of a lipid or lipoprotein in nematodes comprising:
(a) subjecting nematodes that comprise at least one mutation in the clk-1 gene to mutagenesis to produce test nematodes; (b) identifying test nematodes that exhibit a phenotype that is modified as compared to the phenotype of nematodes not subjected to mutagenesis, wherein said phenotype that is modified indicates a modulated level of a lipid or lipoprotein, said phenotype being selected from the group consisting of (i) length of defecation cycle; (ii) rate of germline development relative to rate of soma development; (iii) rate of embryonic development; and (iv) rate of post-embryonic development; and (c) cloning the gene that was mutated in step (a) from a test nematode of step (b).
- 3. A method for identifying a gene that modulates the level of a lipid or lipoprotein in nematodes comprising (a) contacting test nematodes that comprise at least one mutation in the clk-1 gene with a nucleic acid that reduces specifically the level of expression of a nematode gene; and (b) identifying test nematodes that exhibit a phenotype that is modified as compared to the phenotype of nematodes not contacted with said nucleic acid, wherein a modification of said phenotype in said test nematodes indicates that the nematode gene in said test nematodes modulates the level of a lipid or lipoprotein, said phenotype being selected from the group consisting of (i) length of defecation cycle; (ii) rate of germline development relative to rate of soma development; (iii) rate of embryonic development; and (iv) rate of post-embryonic development.
- 4. The method of claim 3, wherein the nucleic acid is selected from the group consisting of an antisense nucleic acid, and a double-stranded RNA molecule.
- 5. The method of claim 4, wherein said contacting comprises feeding nematode with bacteria comprising said nucleic acid.
- 6. A method for selecting nematodes having modulated level of a lipid or lipoprotein comprising:
(a) treating test nematodes to modulate the level of a lipid or lipoprotein; and (b) identifying test nematodes that exhibit a phenotype that is modified as compared to the phenotype of nematodes that has not been treated, wherein a modulated level of a lipid or lipoprotein in said test nematodes is indicated by the phenotype that is modified, said phenotype selected from the group consisting of (i) length of defecation cycle; (ii) rate of germline development relative to rate of soma development; (iii) rate of embryonic development; and (iv) rate of post-embryonic development.
- 7. The method of claim 1 or 6, wherein the test nematodes comprise (a) at least one mutation in the clk-1 gene; (b) at least one mutation in the clk-1 gene and at least one mutation in the dsc-3 gene; or (c) at least one mutation in the clk-1 gene and at least one mutation in the dsc-4 gene.
- 8. The method of claim 1, 2, 3, 4, 5, or 6, wherein the phenotype of said test nematodes in step (b) is modified by (i) a decreased length of defecation cycle; (ii) an increased rate of germline development relative to rate of soma development; (iii) an increased rate of embryonic development; or (iv) an increased rate of post-embryonic development.
- 9. An isolated nucleic acid comprising:
(a) the nucleotide sequence of SEQ ID NO:1 or SEQ ID NO:7; (b) a nucleotide sequence that encodes the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:8 (c) a nucleotide sequence that is at least 80% identical to the nucleotide sequence of SEQ ID NO:1 or SEQ ID NO:7; (d) a nucleotide sequence that is a complement of (a), (b) or (c), with the proviso that the nucleic acid does not comprise genomic sequences that are contiguous to SEQ ID NO:1 or 7, or that are contiguous to a subsequence of SEQ ID NO:1 or 7.
- 10. An isolated nucleic acid that hybridizes under stringent conditions to:
(a) a nucleic acid probe that consists of the nucleotide sequence of SEQ ID NO:1 or SEQ ID NO:7; or (b) a complement of (a); with the proviso that the isolated nucleic acid is not the nucleic acid of one of the clones designated yk357a6, K02D7, H06H21, or Y17G9.
- 11. The nucleic acid of claim 10, wherein the nucleic acid is a double-stranded RNA molecule.
- 12. The nucleic acid of claim 10 wherein the nucleic acid comprises the nucleotide sequence of SEQ ID NO:1 except that the nucleotide residue at nucleotide 354 is a thymine residue and the nucleotide at position 605 is an adenine residue.
- 13. The nucleic acid of claim 10 wherein the nucleic acid comprises a nucleotide sequence that encodes a polypeptide consisting of a fragment of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:8, wherein said polypeptide displays one or more functional activities of the DSC-3 or DSC-4 polypeptide.
- 14. An isolated polypeptide comprising SEQ ID NO:2 or SEQ ID NO:8.
- 15. A fragment of a polypeptide comprising at least 8 consecutive amino acids of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:8.
- 16. A polypeptide comprising an amino acid sequence that has at least 60% identity to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:8.
- 17. The polypeptide of claim 14 wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:2 except that the amino acid residue at position 62 is a phenylalanine residue and the amino acid residue at position 146 is a threonine residue.
- 18. An antibody that immunospecifically binds to a polypeptide comprising the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:8, or a fragment thereof.
- 19. An expression vector comprising the nucleic acid of claim 9.
- 20. A cell comprising a recombinant nucleic acid comprising the nucleic acid of claim 9.
- 21. A transgenic non-human animal comprising a transgene that comprises the nucleic acid of claim 9 or 10.
- 22. The transgenic non-human animal of claim 21, which is a C. elegans nematode.
- 23. A method for making a polypeptide comprising the steps of:
(a) culturing a cell comprising a recombinant MOLL nucleic acid under conditions that allow a MOLL polypeptide to be expressed by said cell; and (b) isolating the MOLL polypeptide.
- 24. A method for identifying a compound that binds a MOLL polypeptide comprising the steps of:
(a) contacting a MOLL polypeptide with a test compound under conditions that allow said test compound to bind said MOLL polypeptide; and (b) detecting binding of said MOLL polypeptide to said test compound.
- 25. A method for preventing or treating atherosclerosis or a dyslipidemia disorder in humans, said method comprising administering to a human in need thereof an amount of a pharmaceutical composition comprising an agonist or an antagonist of a MOLL polypeptide.
- 26. A method for preventing or treating atherosclerosis in humans, said method comprising administering to a human in need thereof an amount of a pharmaceutical composition comprising an clk-1 antagonist.
- 27. The method of claim 23 wherein said MOLL nucleic acid is dsc-3 or dsc-4.
- 28. The method of claim 24 or 25 wherein said MOLL polypeptide is DSC-3 or DSC-4.
- 29. The method of claim 1, 2, 3, or 6, wherein said lipid is cholesterol.
- 30. The method of claim 1, 2, 3, or 6, wherein said lipoprotein is LDL-like lipoprotein.
- 31. The method of claim 1, 2, 3, or 6, wherein a modified phenotype is detected by (i) visual inspection of the phenotype; (ii) the expression profile of one or more indicator genes; (iii) the expression of one or more reporter genes; (iv) a change in the level and/or distribution of a lipid or lipoprotein in the test nematodes.
Parent Case Info
[0001] The present application claims the benefit of U.S. provisional application No. 60/454,925, filed on Mar. 14, 2003, which is incorporated herein by reference in its entirety.
Provisional Applications (1)
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Number |
Date |
Country |
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60454925 |
Mar 2003 |
US |