公开(公告)号：US20070026431A1

公开(公告)日：2007-02-01

申请号：US11441806

申请日：2006-05-26

Applicant: Hiroshi Okabe ,  Frank McCormick ,  Osamu Tetsu

Inventor： Hiroshi Okabe ,  Frank McCormick ,  Osamu Tetsu

IPC: C12Q1/68 ,  G01N33/567 ,  C12Q1/48 ,  C07H21/04 ,  C12P21/06 ,  C12N9/10

CPC classification number: G01N33/5011 ,  C12Q1/485 ,  G01N2333/4739 ,  G01N2500/00

Abstract: The invention features methods and compositions for screening for agents that modulate cellular proliferation, particularly in cells that have elevated cyclin D1 (e.g., cancerous cells), where the methods provide for detection of agents that modulate phosphorylation of cyclin D1 by MAPK and/or detection of agents that modulate ubiquitination of cyclin D1 by FBXW8. The invention also features methods of controlling cellular proliferation, and agents useful in such methods.

Abstract translation: 本发明的特征在于筛选调节细胞增殖的试剂的方法和组合物，特别是在具有升高的细胞周期蛋白D1（例如癌细胞）的细胞中，其中所述方法提供了通过MAPK和/或检测调节细胞周期蛋白D1的磷酸化的试剂 通过FBXW8调节细胞周期蛋白D1泛素化的药剂。 本发明还特征在于控制细胞增殖的方法以及在这些方法中有用的试剂。

公开(公告)号：US06787321B1

公开(公告)日：2004-09-07

申请号：US09687593

申请日：2000-10-13

Applicant: Osamu Tetsu ,  Kenichi Wakita ,  Frank McCormick

Inventor： Osamu Tetsu ,  Kenichi Wakita ,  Frank McCormick

IPC: G01N3353

CPC classification number: C12N15/1055 ,  A61K48/00 ,  A61K48/0058 ,  C12Q1/6897 ,  C12Q2565/201

Abstract: This invention utilizes a two-hybrid system to screen for agents that modulate the ability of a cell to degrade or to accumulate a metabolic product or to selective kill a cell or to selectively express a gene or cDNA in a cell that has a defect in its ability to degrade or to accumulate a metabolic product. One embodiment provides a mammalian cell comprising a nucleic acid encoding a peptide binding domain and an effector gene; a first chimeric protein comprising a nucleic acid binding domain that binds the peptide binding domain attached to the metabolic product or to a ligand that binds to the metabolic product; and a second chimeric protein comprising an expression control protein attached to the metabolic product or to the ligand that binds to the metabolic product such that when the first chimeric protein comprises the metabolic product, the second chimeric protein comprises the ligand and when the first chimeric protein comprises the ligand, the second chimeric protein comprises the metabolic product. The cell is contacted with a test agent and in alteration of expression of the effector gene is detected (if present) where a change in expression of the effector gene in response to the test agent indicates that the test agent modulates the ability of the cell to accumulate or degrade the metabolic product.

Abstract translation: 本发明利用双杂交系统筛选调节细胞降解能力或积累代谢产物或选择性杀死细胞或选择性地在其中具有缺陷的细胞中表达基因或cDNA的试剂 降解或积累代谢产物的能力。 一个实施方案提供了哺乳动物细胞，其包含编码肽结合结构域和效应基因的核酸; 第一嵌合蛋白，其包含结合与所述代谢产物连接的肽结合结构域的核酸结合结构域或与所述代谢产物结合的配体; 以及第二嵌合蛋白，其包含与代谢产物连接的表达控制蛋白或与所述代谢产物结合的配体，使得当所述第一嵌合蛋白包含所述代谢产物时，所述第二嵌合蛋白包含所述配体，并且当所述第一嵌合蛋白 包含配体，第二嵌合蛋白包含代谢产物。 细胞与测试试剂接触，检测效应基因表达的改变（如果存在），其中响应于测试试剂的效应基因的表达变化表明测试试剂调节细胞的能力 积累或降解代谢产物。

公开(公告)号：US07358086B2

公开(公告)日：2008-04-15

申请号：US10896600

申请日：2004-07-21

Applicant: Osamu Tetsu ,  Kenichi Wakita ,  Frank McCormick

Inventor： Osamu Tetsu ,  Kenichi Wakita ,  Frank McCormick

IPC: C12N5/22

CPC classification number: C12N15/1055 ,  A61K48/00 ,  A61K48/0058 ,  C12Q1/6897 ,  C12Q2565/201

Abstract: This invention utilizes a two-hybrid system to screen for agents that modulate the ability of a cell to degrade or to accumulate a metabolic product or to selective kill a cell or to selectively express a gene or cDNA in a cell that has a defect in its ability to degrade or to accumulate a metabolic product. One embodiment provides a mammalian cell comprising a nucleic acid encoding a peptide binding domain and an effector gene; a first chimeric protein comprising a nucleic acid binding domain that binds the peptide binding domain attached to the metabolic product or to a ligand that binds to the metabolic product; and a second chimeric protein comprising an expression control protein attached to the metabolic product or to the ligand that binds to the metabolic product such that when the first chimeric protein comprises the metabolic product, the second chimeric protein comprises the ligand and when the first chimeric protein comprises the ligand, the second chimeric protein comprises the metabolic product. The cell is contacted with a test agent and an alteration of expression of the effector gene is detected (if present) where a change in expression of the effector gene in response to the test agent indicates that the test agent modulates the ability of the cell to accumulate or degrade the metabolic product.

Abstract translation: 本发明利用双杂交系统筛选调节细胞降解能力或积累代谢产物或选择性杀死细胞或选择性地在其中具有缺陷的细胞中表达基因或cDNA的试剂 降解或积累代谢产物的能力。 一个实施方案提供了哺乳动物细胞，其包含编码肽结合结构域和效应基因的核酸; 第一嵌合蛋白，其包含结合与所述代谢产物连接的肽结合结构域的核酸结合结构域或与所述代谢产物结合的配体; 以及第二嵌合蛋白，其包含与代谢产物连接的表达控制蛋白或与所述代谢产物结合的配体，使得当所述第一嵌合蛋白包含所述代谢产物时，所述第二嵌合蛋白包含所述配体，并且当所述第一嵌合蛋白 包含配体，第二嵌合蛋白包含代谢产物。 细胞与测试剂接触，检测效应基因表达的改变（如果存在），其中效应基因响应于测试试剂的表达变化表明测试试剂调节细胞的能力 积累或降解代谢产物。