Methods and reagents for modulating cholesterol levels

Abstract

The invention features ABC1 nucleic acids and polypeptides for the diagnosis and treatment of abnormal cholesterol regulation. The invention also features methods for identifying compounds for modulating cholesterol levels in an animal (e.g., a human).

What is claimed is:  
     
         1 . An isolated ABCA1 polypeptide having ABCA1 biological activity and comprising an amino acid sequence with at least 50% identity to SEQ ID NO: 1.  
     
     
         2 . The isolated ABCA1 polypeptide of  claim 1  wherein said ABCA1 polypeptide is at least 75% pure by weight.  
     
     
         3 . The isolated ABCA1 polypeptide of  claim 1  wherein said ABCA1 polypeptide is at least 90% pure by weight.  
     
     
         4 . The isolated ABCA1 polypeptide of  claim 1  wherein said ABCA1 polypeptide is at least 99% pure by weight.  
     
     
         5 . The isolated ABCA1 polypeptide of  claim 1  wherein said percent identity is at least 85%.  
     
     
         6 . The isolated ABCA1 polypeptide of  claim 1  wherein said percent identity is at least 90%.  
     
     
         7 . The isolated ABCA1 polypeptide of  claim 1  wherein said percent identity is at least 95%.  
     
     
         8 . The isolated ABCA1 polypeptide of  claim 1  wherein said polypeptide comprises the amino acid sequence of SEQ ID NO: 1.  
     
     
         9 . The isolated ABCA1 polypeptide of  claim 1  wherein said ABCA1 polypeptide is a mammalian ABCA1.  
     
     
         10 . The isolated ABCA1 polypeptide of  claim 1  wherein said polypeptide is chicken ABCA1.  
     
     
         11 . The isolated ABCA1 polypeptide of  claim 9  wherein said polypeptide is mouse ABCA1.  
     
     
         12 . The isolated ABCA1 polypeptide of  claim 9  wherein said polypeptide is human ABCA1.  
     
     
         13 . The isolated ABCA1 polypeptide of  claim 1 , wherein said polypeptide comprises amino acids 1 to 60 of SEQ ID NO: 1.  
     
     
         14 . The isolated ABCA1 polypeptide of  claim 1 , wherein said polypeptide comprises amino acids 61 to 2261 of SEQ ID NO: 1.  
     
     
         15 . The isolated ABCA1 polypeptide of  claim 1 , wherein said polypeptide comprises amino acids 1 to 2261 of SEQ ID NO: 1.  
     
     
         16 . The isolated ABCA1 polypeptide of  claim 1  wherein said polypeptide is a recombinant polypeptide.  
     
     
         17 . The isolated ABCA1 polypeptide of  claim 1  wherein said polypeptide is derived from a natural source.  
     
     
         18 . The isolated ABCA1 polypeptide of  claim 1  wherein said biological activity is lipid transport across a membrane.  
     
     
         19 . The isolated ABCA1 polypeptide of  claim 19  wherein said lipid is member selected from the group consisting of cholesterol and phospholipid.  
     
     
         20 . The isolated ABCA1 polypeptide of  claim 1  wherein said biological activity is transport of an ion across a membrane.  
     
     
         21 . The isolated ABCA1 polypeptide of  claim 1  wherein said biological activity is transport of interleukin-1 across a membrane.  
     
     
         22 . An isolated ABCA1 polypeptide comprising amino acids 1 to 60 of SEQ ID NO: 1.  
     
     
         23 . An isolated ABCA1 polypeptide comprising amino acids 61 to 2261 of SEQ ID NO: 1.  
     
     
         24 . An isolated ABCA1 polypeptide comprising amino acids 1 to 2261 of SEQ ID NO: 1.  
     
     
         25 . An isolated polynucleotide comprising a nucleotide sequence having at least 50% identity to SEQ ID NO: 1 and that encodes an ABCA1 polypeptide having ABCA1 biological activity.  
     
     
         26 . The isolated polynucleotide of  claim 25  wherein said polynucleotide is at least 75% pure by weight.  
     
     
         27 . The isolated polynucleotide of  claim 25  wherein said polynucleotide purity is at least 90% pure by weight.  
     
     
         28 . The isolated polynucleotide of  claim 25  wherein said polynucleotide is at least 99% pure by weight.  
     
     
         29 . The isolated polynucleotide of  claim 25  wherein said polynucleotide is at least 99% pure by weight.  
     
     
         30 . The isolated polynucleotide of  claim 25  wherein said percent identity is at least 85%.  
     
     
         31 . The isolated polynucleotide of  claim 25  wherein said percent identity is at least 90%.  
     
     
         32 . The isolated polynucleotide of  claim 25  wherein said percent identity is at least 95%.  
     
     
         33 . The isolated polynucleotide of  claim 25  wherein said polynucleotide acid comprises the nucleotide sequence of SEQ ID NO: 2.  
     
     
         34 . The isolated polynucleotide of  claim 25  wherein said polynucleotide acid hybridizes at high stringency conditions to nucleotides 75 to 254 of SEQ ID NO: 2.  
     
     
         35 . The isolated polynucleotide of  claim 25  wherein said polynucleotide comprises nucleotides 75 to 254 of SEQ ID NO: 2.  
     
     
         36 . The isolated polynucleotide of  claim 25  wherein said polynucleotide comprises nucleotides 255 to 6858 of SEQ ID NO: 2.  
     
     
         37 . The isolated polynucleotide of  claim 25  wherein said polynucleotide comprises nucleotides 75 to 6857 of SEQ ID NO: 2.  
     
     
         38 . The isolated polynucleotide of  claim 25  wherein said polynucleotide comprises nucleotides 75 to 254 of SEQ ID NO: 2.  
     
     
         39 . The isolated polynucleotide of  claim 25  wherein said polynucleotide comprises nucleotides 75 to 6857 of SEQ ID NO: 2.  
     
     
         40 . The isolated polynucleotide of  claim 25  that comprises nucleotides 255 to 6857 of SEQ ID NO: 2.  
     
     
         41 . A substantially pure polynucleotide encoding the ABCA1 polypeptide of  claim 1 .  
     
     
         42 . An expression vector comprising the polynucleotide of  claim 25 .  
     
     
         43 . An expression vector comprising the polynucleotide of  claim 41 .  
     
     
         44 . A recombinant cell genetically engineered to express the polynucleotide of  claim 25 .  
     
     
         45 . The recombinant cell of  claim 44  wherein said cell does not express said polynucleotide absent said genetic engineering.  
     
     
         46 . A recombinant cell expressing the polynucleotide of  claim 41 .  
     
     
         47 . The recombinant cell of  claim 46  wherein said cell does not express said polynucleotide absent said genetic engineering.  
     
     
         48 . A method of making an ABCA1 polypeptide comprising expressing said polypeptide from the recombinant cell of  claim 44 .  
     
     
         49 . A method of making an ABCA1 polypeptide comprising expressing said polypeptide from the recombinant cell of  claim 46 .  
     
     
         50 . A transgenic non-human animal that expresses the polynucleotide of  claim 25 .  
     
     
         51 . A transgenic non-human animal that expresses the polynucleotide of  claim 41 .  
     
     
         52 . A substantially pure polynucleotide comprising at least thirty consecutive nucleotides contained in nucleotides 7015-7860 of SEQ ID NO: 2.  
     
     
         53 . The substantially pure polynucleotide of  claim 52  wherein said polynucleotide comprises nucleotides 7015-7860 of SEQ ID NO: 2.  
     
     
         54 . A substantially pure polynucleotide that hybridizes at high stringency to a probe comprising nucleotides 7015-7860 of SEQ ID NO: 2.  
     
     
         55 . An oligonucleotide comprising nucleotides 75 to 254 of SEQ ID NO: 2.  
     
     
         56 . The polynucleotide of  claim 25  wherein said polynucleotide is a cDNA.  
     
     
         57 . The polynucleotide of  claim 41  wherein said polynucleotide is a cDNA.  
     
     
         58 . A cell engineered to contain in its membrane the ABCA1 polypeptide of  claim 1  wherein said cell does not contain said polypeptide absent said engineering.  
     
     
         59 . The cell of  claim 58  wherein said engineering is genetic engineering.  
     
     
         60 . The cell of  claim 58  wherein said cell is a recombinant cell.  
     
     
         61 . The cell of  claim 58  wherein said cell is a mammalian cell.  
     
     
         62 . The cell of  claim 61  wherein said mammalian cell is a human cell.  
     
     
         63 . A cell engineered to contain in its membrane the ABCA1 polypeptide of  claim 22  wherein said cell does not contain said polypeptide absent said engineering.  
     
     
         64 . A cell engineered to contain in its membrane the ABCA1 polypeptide of  claim 23  wherein said cell does not contain said polypeptide absent said engineering.  
     
     
         65 . A cell engineered to contain in its membrane the ABCA1 polypeptide of  claim 24  wherein said cell does not contain said polypeptide absent said engineering.