公开(公告)号：US20090191581A1

公开(公告)日：2009-07-30

申请号：US12245546

申请日：2008-10-03

申请人： Ronald T. Raines ,  Stephen M. Fuchs

发明人： Ronald T. Raines ,  Stephen M. Fuchs

IPC分类号： C12Q1/37 ,  C07K19/00

CPC分类号： C12Q1/37 ,  C07K14/43595 ,  G01N33/5005

摘要： This invention relates to methods and compositions for designing novel fluorescent proteins, preferably to a green fluorescent proteins (GFP). The engineered GFPs are modified by substituting negatively charged amino acids with positively charged amino acids on the exterior of the protein making the protein cell permeable. The ability of the engineered fluorescent proteins to permeate cells obviates the need for transfections, allowing these novel proteins to be used in numerous biological applications.

摘要翻译： 本发明涉及用于设计新的荧光蛋白，优选绿色荧光蛋白（GFP）的方法和组合物。 通过在蛋白质外部用带正电荷的氨基酸取代带负电荷的氨基酸来修饰工程化的GFP，使得蛋白质细胞是可渗透的。 工程化荧光蛋白渗透细胞的能力消除了转染的需要，允许这些新型蛋白质用于许多生物应用。

公开(公告)号：US08834918B2

公开(公告)日：2014-09-16

申请号：US12017953

申请日：2008-01-22

申请人： David M. Lynn ,  Ronald T. Raines ,  Christopher M. Jewell ,  Stephen M. Fuchs ,  Ryan M. Flessner

发明人： David M. Lynn ,  Ronald T. Raines ,  Christopher M. Jewell ,  Stephen M. Fuchs ,  Ryan M. Flessner

IPC分类号： A61L15/16 ,  A61K9/70 ,  A61K47/48

CPC分类号： A61K9/7007 ,  A61K47/645

摘要： A composition for delivery of a molecule into a cell is provided. The composition includes a protein transduction domain that is conjugated to the molecule which is incorporated into a multilayered film. Preferably, the protein transduction domain is a cationic protein transduction domain. More preferably, the cationic protein transduction domain is nonaarginine, and the multilayered film includes polyelectrolyte multilayers. When the composition is presented to a cell, the multilayered film dissolves or erodes in physiological media, and the molecule is delivered into the cell.

摘要翻译： 提供了将分子递送到细胞中的组合物。 该组合物包括与分子结合的蛋白转导结构域，其结合到多层膜中。 优选地，蛋白质转导结构域是阳离子蛋白转导结构域。 更优选地，阳离子蛋白转导结构域是非精氨酸，多层膜包括聚电解质多层。 当将组合物提供给细胞时，多层膜在生理介质中溶解或侵蚀，并将分子递送至细胞中。

公开(公告)号：US07452973B2

公开(公告)日：2008-11-18

申请号：US11593664

申请日：2006-11-07

申请人： Ronald T. Raines ,  Stephen M. Fuchs

发明人： Ronald T. Raines ,  Stephen M. Fuchs

IPC分类号： C07K14/00 ,  C12N15/12 ,  C12N15/63 ,  C07H21/04

CPC分类号： C12Q1/37 ,  C07K14/43595 ,  G01N33/5005

摘要： This invention relates to methods and compositions for designing novel fluorescent proteins, preferably to a green fluorescent proteins (GFP). The engineered GFPs are modified by substituting negatively charged amino acids with positively charged amino acids on the exterior of the protein making the protein cell permeable. The ability of the engineered fluorescent proteins to permeate cells obviates the need for transfections, allowing these novel proteins to be used in numerous biological applications.

摘要翻译： 本发明涉及用于设计新的荧光蛋白，优选绿色荧光蛋白（GFP）的方法和组合物。 通过在蛋白质外部用带正电荷的氨基酸取代带负电荷的氨基酸来修饰工程化的GFP，使得蛋白质细胞是可渗透的。 工程化荧光蛋白渗透细胞的能力消除了转染的需要，允许这些新型蛋白质用于许多生物应用。

公开(公告)号：US07973132B2

公开(公告)日：2011-07-05

申请号：US12245546

申请日：2008-10-03

申请人： Ronald T. Raines ,  Stephen M. Fuchs

发明人： Ronald T. Raines ,  Stephen M. Fuchs

IPC分类号： C07K14/00 ,  C12N15/12 ,  C12N15/63 ,  C07H21/04

CPC分类号： C12Q1/37 ,  C07K14/43595 ,  G01N33/5005

摘要： This invention relates to methods and compositions for designing novel fluorescent proteins, preferably to a green fluorescent proteins (GFP). The engineered GFPs are modified by substituting negatively charged amino acids with positively charged amino acids on the exterior of the protein making the protein cell permeable. The ability of the engineered fluorescent proteins to permeate cells obviates the need for transfections, allowing these novel proteins to be used in numerous biological applications.

摘要翻译： 本发明涉及用于设计新的荧光蛋白，优选绿色荧光蛋白（GFP）的方法和组合物。 通过在蛋白质外部用带正电荷的氨基酸取代带负电荷的氨基酸来修饰工程化的GFP，使得蛋白质细胞是可渗透的。 工程化荧光蛋白渗透细胞的能力消除了转染的需要，允许这些新型蛋白质用于许多生物应用。

公开(公告)号：US07098016B2

公开(公告)日：2006-08-29

申请号：US10461713

申请日：2003-06-13

申请人： Ronald T. Raines ,  Parit Plainkum ,  Stephen M. Fuchs

发明人： Ronald T. Raines ,  Parit Plainkum ,  Stephen M. Fuchs

IPC分类号： C12N9/22

CPC分类号： C12N9/22

摘要： An enzyme is re-engineered to be a zymogen, an enzyme precursor which is converted into an enzyme by protease cleavage. In the example described here, an RNase A enzyme is converted into a zymogen by adding to the enzyme a bridge of amino acids linking the amino and carboxyl termini of the enzyme. The bridge has built in it a protease cleavage site for a specific protease, for example the protease plasmepsin II, produced by the malaria parasite. Since RNase A can be made cytotoxic, this permits a cytotoxic enzyme to be made in the form of a zymogen that becomes active only when it is acted on by a protease only present in a particular target cell such as a pathogen.

摘要翻译： 酶被重新设计为酶原，酶原，其通过蛋白酶切割转化成酶。 在本文所述的实施例中，通过向酶加入连接酶的氨基和羧基末端的氨基酸桥，将RNase A酶转化为酵素。 该桥已经建立了特定蛋白酶的蛋白酶切割位点，例如由疟原虫产生的蛋白酶等质粒蛋白酶II。 由于RNA酶A可以被制成细胞毒性，因此允许细胞毒素酶以酶原的形式制备，只有当其被存在于特定靶细胞例如病原体中的蛋白酶作用时才变成活性。