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    Peptides whose uptake by cells is controllable
    1.
    发明申请
    Peptides whose uptake by cells is controllable 有权
    细胞摄取的肽是可控的

    公开(公告)号：US20060041105A1

    公开(公告)日：2006-02-23

    申请号：US11133804

    申请日：2005-05-19

    申请人： Tao Jiang ,  Emilia Olson ,  Michael Whitney ,  Roger Tsien

    发明人： Tao Jiang ,  Emilia Olson ,  Michael Whitney ,  Roger Tsien

    IPC分类号： A61K38/12

    CPC分类号： C07K7/08 ,  A61K38/00 ,  A61K41/0095 ,  A61K47/64 ,  A61K47/645 ,  A61K47/65 ,  A61K49/0032 ,  A61K49/0043 ,  A61K49/0056 ,  A61K49/085 ,  A61K49/14 ,  A61K49/146 ,  A61K51/088 ,  C07K14/00 ,  C07K14/4728

    摘要： A generic structure for the peptides of the present invention includes A-X-B-C, where C is a cargo moiety, the B portion includes basic amino acids, X is a cleavable linker sequence, and the A portion includes acidic amino acids. The intact structure is not significantly taken up by cells; however, upon extracellular cleavage of X, the B-C portion is taken up, delivering the cargo to targeted cells. Cargo may be, for example, a contrast agent for diagnostic imaging, a chemotherapeutic drug, or a radiation-sensitizer for therapy. X may be cleaved extracellularly or intracellularly. The molecules of the present inventnon may be linear, cyclic, branched, or have a mixed structure.

    摘要翻译： 本发明的肽的一般结构包括A-X-B-C，其中C是货物部分，B部分包括碱性氨基酸，X是可切割的接头序列，A部分包括酸性氨基酸。 完整结构不被细胞所占据; 然而，在X的细胞外裂解时，B-C部分被吸收，将货物运送到靶细胞。 货物可以是例如用于诊断成像的对比剂，化学治疗药物或用于治疗的放射线敏化剂。 X可能在细胞外或细胞内裂解。 本发明的分子可以是直链，环状，支链或具有混合结构。

    Peptides whose uptake by cells is controllable
    3.
    发明授权
    Peptides whose uptake by cells is controllable 有权
    细胞摄取的肽是可控的

    公开(公告)号：US07985401B2

    公开(公告)日：2011-07-26

    申请号：US11133804

    申请日：2005-05-19

    申请人： Tao Jiang ,  Emilia S. Olson ,  Michael Whitney ,  Roger Tsien

    发明人： Tao Jiang ,  Emilia S. Olson ,  Michael Whitney ,  Roger Tsien

    IPC分类号： A61M36/14 ,  A61K51/00

    CPC分类号： C07K7/08 ,  A61K38/00 ,  A61K41/0095 ,  A61K47/64 ,  A61K47/645 ,  A61K47/65 ,  A61K49/0032 ,  A61K49/0043 ,  A61K49/0056 ,  A61K49/085 ,  A61K49/14 ,  A61K49/146 ,  A61K51/088 ,  C07K14/00 ,  C07K14/4728

    摘要： A generic structure for the peptides of the present invention includes A-X-B-C, where C is a cargo moiety, the B portion includes basic amino acids, X is a cleavable linker sequence, and the A portion includes acidic amino acids. The intact structure is not significantly taken up by cells; however, upon extracellular cleavage of X, the B-C portion is taken up, delivering the cargo to targeted cells. Cargo may be, for example, a contrast agent for diagnostic imaging, a chemotherapeutic drug, or a radiation-sensitizer for therapy. X may be cleaved extracellularly or intracellularly. The molecules of the present invention may be linear, cyclic, branched, or have a mixed structure.

    摘要翻译： 本发明的肽的一般结构包括A-X-B-C，其中C是货物部分，B部分包括碱性氨基酸，X是可切割的接头序列，A部分包括酸性氨基酸。 完整结构不被细胞所占据; 然而，在X的细胞外裂解时，B-C部分被吸收，将货物运送到靶细胞。 货物可以是例如用于诊断成像的对比剂，化学治疗药物或用于治疗的放射线敏化剂。 X可能在细胞外或细胞内裂解。 本发明的分子可以是直链，环状，支链或具有混合结构。

    Methods of protein destabilization and uses thereof
    7.
    发明申请
    Methods of protein destabilization and uses thereof 有权
    蛋白质不稳定的方法及其用途

    公开(公告)号：US20080227129A1

    公开(公告)日：2008-09-18

    申请号：US11821562

    申请日：2007-06-22

    申请人： Jeffrey Stack ,  Michael Whitney ,  Andrew B. Cubitt ,  Brian Pollok

    发明人： Jeffrey Stack ,  Michael Whitney ,  Andrew B. Cubitt ,  Brian Pollok

    IPC分类号： C12Q1/66 ,  C12Q1/02 ,  C12Q1/37 ,  C12Q1/48 ,  C12N1/16 ,  C07K14/00 ,  C12N5/06 ,  C12N5/00 ,  C12Q1/42 ,  C12Q1/34

    CPC分类号： C12Q1/485

    摘要： This invention is directed towards methods of destabilizing proteins in living cells, and their use for the development of novel assays. In one embodiment, the invention comprises the use of non-cleavable multimerized ubiquitin fusion proteins to destabilize a target protein, such as a reporter moiety. In one aspect of this method the constructs also comprises a linker that operatively couples the reporter moiety to the multimerized ubiquitin fusion protein. In this embodiment, enzymatic modification of the linker results in a modulation of the coupling of the reporter protein to the multimerized ubiquitin domains resulting in a change in the stability of the reporter moiety. The level of the reporter moiety in the cell can then be used as a measure of the enzymatic activity in the cell. In another embodiment the invention provides for a generalized way of coordinately regulating the cellular concentration of a plurality of target proteins. In one aspect of this method, the target proteins are operatively coupled to a ubiquitin fusion protein via a linker containing a protease cleavage site. Cleavage of the linker by a protease results in uncoupling of the target protein from the multimerized ubiquitin construct, and results in an increase in the stability and concentration of the target protein.

    摘要翻译： 本发明涉及使活细胞中的蛋白质失稳的方法及其用于开发新的测定方法。 在一个实施方案中，本发明包括使用不可切割的多聚化泛素融合蛋白来使目标蛋白如报道分子部分不稳定。 在该方法的一个方面，构建体还包含使报道部分与多聚化泛素融合蛋白可操作地连接的接头。 在该实施方案中，接头的酶修饰导致报道蛋白与多聚泛素结构域的偶联的调节，导致报道部分稳定性的变化。 然后可以将细胞中的报告物部分的水平用作细胞中酶活性的量度。 在另一个实施方案中，本发明提供了协调调节多种靶蛋白的细胞浓度的一般化方式。 在该方法的一个方面，靶蛋白通过含有蛋白酶切割位点的接头与泛素融合蛋白可操作地偶联。 通过蛋白酶切割接头导致靶蛋白与多聚化泛素构建体解偶联，并导致靶蛋白的稳定性和浓度增加。

    Methods of Protein Destabilization and Uses Thereof
    8.
    发明申请
    Methods of Protein Destabilization and Uses Thereof 审中-公开
    蛋白质不稳定的方法及其用途

    公开(公告)号：US20110191873A1

    公开(公告)日：2011-08-04

    申请号：US12938179

    申请日：2010-11-02

    申请人： Jeffrey Stack ,  Michael Whitney ,  Andrew B. Cubitt ,  Brian Pollok

    发明人： Jeffrey Stack ,  Michael Whitney ,  Andrew B. Cubitt ,  Brian Pollok

    IPC分类号： A01K67/00 ,  C12N5/10 ,  C07H21/04 ,  C12N1/00 ,  C07K19/00 ,  A01H5/00 ,  C12Q1/02 ,  C12Q1/37 ,  C12Q1/42 ,  C12Q1/48 ,  C12Q1/66 ,  C12N5/07 ,  C12N5/071 ,  C12N5/04

    CPC分类号： C12Q1/485

    摘要： This invention is directed towards methods of destabilizing proteins in living cells, and their use for the development of novel assays. In one embodiment, the invention comprises the use of non-cleavable multimerized ubiquitin fusion proteins to destabilize a target protein, such as a reporter moiety. In one aspect of this method the constructs also comprises a linker that operatively couples the reporter moiety to the multimerized ubiquitin fusion protein. In this embodiment, enzymatic modification of the linker results in a modulation of the coupling of the reporter protein to the multimerized ubiquitin domains resulting in a change in the stability of the reporter moiety. The level of the reporter moiety in the cell can then be used as a measure of the enzymatic activity in the cell. In another embodiment the invention provides for a generalized way of coordinately regulating the cellular concentration of a plurality of target proteins. In one aspect of this method, the target proteins are operatively coupled to a ubiquitin fusion protein via a linker containing a protease cleavage site. Cleavage of the linker by a protease results in uncoupling of the target protein from the multimerized ubiquitin construct, and results in an increase in the stability and concentration of the target protein.

    摘要翻译： 本发明涉及使活细胞中的蛋白质失稳的方法及其用于开发新的测定方法。 在一个实施方案中，本发明包括使用不可切割的多聚化泛素融合蛋白来使目标蛋白如报道分子部分不稳定。 在该方法的一个方面，构建体还包含使报道部分与多聚化泛素融合蛋白可操作地连接的连接体。 在该实施方案中，接头的酶修饰导致报道蛋白与多聚泛素结构域的偶联的调节，导致报道部分稳定性的变化。 然后可以将细胞中的报告物部分的水平用作细胞中酶活性的量度。 在另一个实施方案中，本发明提供了协调调节多种靶蛋白的细胞浓度的一般化方式。 在该方法的一个方面，靶蛋白通过含有蛋白酶切割位点的接头与泛素融合蛋白可操作地偶联。 通过蛋白酶切割接头导致靶蛋白与多聚化泛素构建体解偶联，并导致靶蛋白的稳定性和浓度增加。

    Methods of protein destabilization and uses thereof
    9.
    发明授权
    Methods of protein destabilization and uses thereof 有权
    蛋白质不稳定的方法及其用途

    公开(公告)号：US07262005B1

    公开(公告)日：2007-08-28

    申请号：US09498098

    申请日：2000-02-04

    申请人： Jeffrey Stack ,  Michael Whitney ,  Andrew B. Cubitt ,  Brian Pollok

    发明人： Jeffrey Stack ,  Michael Whitney ,  Andrew B. Cubitt ,  Brian Pollok

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/485

    摘要： This invention is directed towards methods of destabilizing proteins in living cells, and their use for the development of novel assays. In one embodiment, the invention comprises the use of non-cleavable multimerized ubiquitin fusion proteins to destabilize a target protein, such as a reporter moiety. In one aspect of this method the constructs also comprises a linker that operatively couples the reporter moiety to the multimerized ubiquitin fusion protein. In this embodiment, enzymatic modification of the linker results in a modulation of the coupling of the reporter protein to the multimerized ubiquitin domains resulting in a change in the stability of the reporter moiety. The level of the reporter moiety in the cell can then be used as a measure of the enzymatic activity in the cell. In another embodiment the invention provides for a generalized way of coordinately regulating the cellular concentration of a plurality of target proteins. In one aspect of this method, the target proteins are operatively coupled to a ubiquitin fusion protein via a linker containing a protease cleavage site. Cleavage of the linker by a protease results in uncoupling of the target protein from the multimerized ubiquitin construct, and results in an increase in the stability and concentration of the target protein.

    摘要翻译： 本发明涉及使活细胞中的蛋白质失稳的方法及其用于开发新的测定方法。 在一个实施方案中，本发明包括使用不可切割的多聚化泛素融合蛋白来使目标蛋白如报道分子部分不稳定。 在该方法的一个方面，构建体还包含使报道部分与多聚化泛素融合蛋白可操作地连接的接头。 在该实施方案中，接头的酶修饰导致报道蛋白与多聚泛素结构域的偶联的调节，导致报道部分稳定性的变化。 然后可以将细胞中的报告物部分的水平用作细胞中酶活性的量度。 在另一个实施方案中，本发明提供了协调调节多种靶蛋白的细胞浓度的一般化方式。 在该方法的一个方面，靶蛋白通过含有蛋白酶切割位点的接头与泛素融合蛋白可操作地偶联。 通过蛋白酶切割接头导致靶蛋白与多聚化泛素构建体解偶联，并导致靶蛋白的稳定性和浓度增加。