Nucleotide and protein sequences of lats genes and methods based thereon
AssigneeYale University
InventorsTian Xu, Wufan Tao, Weiyi Wang, Sheng Zhang, Wan Yu
Life of this patent
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Abstract
The present invention relates to a tumor suppressor gene, termed large tumor suppressor (lats). The invention provides nucleotide sequences of lats genes, and amino acid sequences of their encoded proteins, as well as derivatives (e.g., fragments) and analogs thereof. Diagnostic methods and pharmaceutical compositions are also provided. The invention further relates to treatment or prevention of disease or disorder involving cell proliferation by administering compounds that promote lats activity (e.g., lats, an agonist of lats; nucleic acids that encode lats).
Claims103
A purified lats protein.
A purified protein encoded by a nucleic acid hybridizable to the lats DNA sequence in plasmid PBS(KS)-h-lats as deposited with the ATCC and assigned accession number 69769.
A purified protein encoded by a nucleic acid hybridizable to a DNA having a sequence consisting of the coding region of SEQ ID NO:7.
A purified fragment of a lats protein comprising a domain of the lats protein selected from the group consisting of a lats C-terminal domain 3 (LCD3), lats C-terminal domain 2 (LCD2), lats C-terminal domain 1 (LCD1), kinase domain, a kinase subdomain, lats flanking domain (LFD), lats split domain 1 (LSD1), lats split domain 2 (LSD2), SH3-binding domain, and opa repeat domain.
A protein comprising an amino acid sequence that has at least 60% identity to a domain of a lats protein, in which the percentage identity is determined over an amino acid sequence of identical size to the domain.
A protein comprising an amino acid sequence that has at least 90% identity to a domain of a lats protein, in which the percentage identity is determined over an amino acid sequence of identical size to the domain.
A chimeric protein comprising a fragment of a lats protein consisting of at least 6 amino acids fused via a covalent bond to an amino acid sequence of a second protein, in which the second protein is not a lats protein.
An antibody which is capable of binding a lats protein.
An isolated nucleic acid comprising a nucleotide sequence encoding a lats protein.
An isolated nucleic acid comprising the lats coding sequence contained in plasmid pBS(KS)-h-lats as deposited with the ATCC and assigned accession number 69769.
An isolated nucleic acid hybridizable to the lats DNA sequence in plasmid pBS(KS)-h-lats as deposited with the ATCC and assigned accession number 69769.
An isolated nucleic acid hybridizable to a DNA having a sequence consisting of the coding region of SEQ ID NO:7.
An isolated nucleic acid comprising a fragment of a lats gene consisting of at least 8 nucleotides.
An isolated nucleic acid comprising a nucleotide sequence encoding a fragment of a lats protein that displays one or more functional activities of the lats protein.
An isolated nucleic acid comprising a nucleotide sequence encoding a protein, said protein comprising the amino acid sequence of SEQ ID NO:4.
A pharmaceutical composition comprising a therapeutically effective amount of a lats protein;
- and a pharmaceutically acceptable carrier.
A pharmaceutical composition comprising a therapeutically effective amount of an antibody that immunospecifically binds to a lats protein;
- and a pharmaceutically acceptable carrier.
A pharmaceutical composition comprising a therapeutically effective amount of a fragment or derivative of an antibody that immunospecifically binds to a lats protein, said fragment or derivative containing the binding domain of the antibody;
- and a pharmaceutically acceptable carrier.
A method of treating or preventing a disease or disorder involving cell overproliferation in a subject comprising administering to a subject in which such treatment or prevention is desired a therapeutically effective amount of a molecule that promotes lats function.
A method of treating or preventing a disease or disorder involving a deficiency in cell proliferation or in which cell proliferation is desirable for treatment or prevention in a subject comprising administering to a subject in which such treatment or prevention is desired a therapeutically effective amount of a molecule that inhibits lats function.
An isolated oligonucleotide consisting of at least six nucleotides, and comprising a sequence complementary to at least a portion of an RNA transcript of a lats gene, which oligonucleotide is hybridizable to the RNA transcript under moderately stringent conditions.
A method of diagnosing a disease or disorder characterized by an aberrant level of lats RNA or protein in a subject, comprising measuring the level of lats RNA or protein in a sample derived from the subject, in which an increase or decrease in the level of lats RNA or protein, relative to the level of lats RNA or protein found in an analogous sample not having the disease or disorder indicates the presence of the disease or disorder in the subject.
A method of diagnosing or screening for the presence of or a predisposition for developing a disease or disorder involving cell overproliferation in a subject comprising measuring the level of lats protein, lats RNA or lats functional activity in a sample derived from the subject, in which a decrease in the level of lats protein, lats RNA, or lats functional activity in the sample, relative to the level of lats protein, lats RNA, or lats functional activity found in an analogous sample not having the disease or disorder or a predisposition for developing the disease or disorder, indicates the presence of the disease or disorder or a predisposition for developing the disease or disorder.
A method of diagnosing or screening for the presence of or a predisposition for developing a disease or disorder involving cell overproliferation in a subject comprising detecting one or more mutations in lats DNA, RNA or protein derived from the subject in which the presence of said one or more mutations indicates the presence of the disease or disorder or a predisposition for developing the disease or disorder.
A method of diagnosing or screening for the presence of or a predisposition for developing a disease or disorder involving a deficiency in cell proliferation or in which cell proliferation is desirable for treatment or prevention in a subject comprising measuring the level of lats protein, lats RNA or lats functional activity in a sample derived from the subject, in which an increase in the level of lats protein, lats RNA, or lats functional activity in the sample, relative to the level of lats protein, lats RNA, or lats functional activity found in an analogous sample not having the disease or disorder or a predisposition for developing the disease or disorder, indicates the presence of the disease or disorder or a predisposition for developing the disease or disorder.
A kit comprising in one or more containers a molecule selected from the group consisting of an anti-lats antibody, a nucleic acid probe capable of hybridizing to a lats RNA, or a pair of nucleic acid primers capable of priming amplification of at least a portion of a lats nucleic acid.
A kit comprising in a container a therapeutically effective amount of a lats protein.
A method of increasing cell growth in animals or plants comprising inhibiting lats expression or activity in said animals or plants.
A method of identifying a molecule that specifically binds to a ligand selected from the group consisting of a lats protein, a fragment of a lats protein comprising a domain of the protein, and a nucleic acid encoding the protein or fragment, comprising
- (a) contacting said ligand with a plurality of molecules under conditions conducive to binding between said ligand and the molecules;
- and
- (b) identifying a molecule within said plurality that specifically binds to said ligand.
A recombinant non-human animal or plant that is the product of a process comprising introducing a nucleic acid encoding at least a domain of a lats protein into the plant or animal.
A recombinant plant containing and capable of expressing a lats antisense nucleic acid.
A recombinant non-human animal or plant in which a lats gene has been inactivated by a method comprising introducing a nucleic acid into the plant or animal or an ancestor thereof, which nucleic acid comprises a non-lats sequence flanked by lats genomic sequences that promote homologous recombination.
A method of identifying a tumor suppressor gene comprising (a) identifying an overproliferation phenotype in a genetic mosaic;
- and (b) isolating a gene that is mutated in cells exhibiting said overproliferation phenotype.
A non-human mammal comprising (a) a nucleic acid sequence encoding a recombinase operably linked to a promoter;
- and (b) intrachromosomal insertions into the homologous arms of both of a set of parental chromosomes, of a target site at which the recombinase can promote recombination.
A method for identifying a gene with an identifiable mutant phenotype comprising:
- (a) identifying a mutant phenotype in a genetic mosaic animal, said genetic mosaic animal having been produced by a method comprising recombinantly expressing a recombinase within a cell of the animal so as to promote recombination at intrachromosomally inserted target sites on the homologous arms of both of a set of parental chromosomes;
- and
- (b) isolating a gene that is mutated in cells exhibiting said mutant phenotype.
A method for identifying a gene with an identifiable mutant phenotype comprising:
- (a) identifying a mutant phenotype in a cultured cell, said cultured cell having been produced by a method comprising recombinantly expressing a recombinase within said cultured cell so as to promote recombination at intrachromosomally inserted target sites on the homologous arms of both of a set of parental chromosomes;
- and
- (b) isolating a gene that is mutated in cells exhibiting said mutant phenotype.
A method of inhibiting cellular senescence in a subject comprising administering to a subject in which such inhibition is desired an amount of a molecule that inhibits lats function, effective to inhibit cellular senescence.
A method of inhibiting cellular senescence in cells in vitro comprising contacting cells in vitro with an amount of a molecule that inhibits lats function, effective to inhibit cellular senescence.
Dependencies are inferred from the claim text.