公开(公告)号：US20050123909A1

公开(公告)日：2005-06-09

申请号：US09229229

申请日：1999-01-12

申请人： GEOFFREY M. WAHL ,  NORIAKI SHIMIZU ,  TERU KANDA ,  H. MICHAEL SHEPARD

发明人： GEOFFREY M. WAHL ,  NORIAKI SHIMIZU ,  TERU KANDA ,  H. MICHAEL SHEPARD

IPC分类号： A61K45/00 ,  A61K48/00 ,  A61K49/00 ,  A61P35/00 ,  A61P43/00 ,  C07K14/00 ,  C12N15/09 ,  C12Q1/02 ,  C12Q1/68 ,  G01N33/15 ,  G01N33/50 ,  G01N33/52 ,  G01N33/574 ,  G01N33/58 ,  G01N33/68

CPC分类号： C12Q1/68 ,  G01N33/5011 ,  G01N33/6875 ,  G01N2333/43595

摘要： This invention provides methods by which test substances can be screened for their ability to inhibit, enhance or eliminate double minute (DM) or extrachromosomal DNA by micronucleation in cells. This invention also provides a method for inducing maturation or death of a cell having the capacity to generate micronuclei. It also provides a method of treating a disease in a subject, the cells correlated with the disease having DM and extrachromosomal DNA as well as the capacity to generate micronuclei to capture them. Further provided is a method of detecting chromosomal and extrachromosomal DNA in a cell.

摘要翻译： 本发明提供了通过细胞中微核来筛选测试物质以抑制，增强或消除双分钟（DM）或染色体外DNA的能力的方法。 本发明还提供了诱导具有产生微核能力的细胞的成熟或死亡的方法。 它还提供了治疗受试者疾病的方法，与具有DM和染色体外DNA的疾病相关的细胞以及产生微核以捕获它们的能力。 还提供了检测细胞中染色体和染色体外DNA的方法。

公开(公告)号：US4833251A

公开(公告)日：1989-05-23

申请号：US122496

申请日：1987-11-16

申请人： Gary F. Musso ,  Soumitra Ghosh ,  Leslie E. Orgel ,  Geoffrey M. Wahl ,  Emil T. Kaiser

发明人： Gary F. Musso ,  Soumitra Ghosh ,  Leslie E. Orgel ,  Geoffrey M. Wahl ,  Emil T. Kaiser

IPC分类号： C07F7/18 ,  C07H21/00 ,  C12Q1/68

CPC分类号： C07H21/00 ,  C07F7/1852 ,  C07F7/1856 ,  C12Q1/6816

摘要： Nucleic acid hybridization probes are provided which comprise an N.sup.4 -(substituted amino)cytosine moiety, wherein the substituted amino group comprises a tag moiety, whereby the probe is detected. Methods of preparing probes of the invention, intermediates used in such methods, and methods of using the probes of the invention in hybridization assays are also provided. Typical tag moieties employed with the probes of the invention are biotinyl, aminothiadiazole and fluorescein derivatives, connected to N.sup.4 -amino groups of modified cytosines of the probe through linker moieties. Probes tagged with biotin are typically detected by binding to the biotinyl moieties, through a streptavidin or avidin molecule, a reporter group which includes streptavidin or avidin and then detecting a signal due to the reporter group. Probes tagged with aminothiadiazole derivatives are typically detected by essentially the same method as those tagged with biotinyl but employing as reporter group one which binds to the derivative through a carbonic anhydrase molecule. Probes tagged with fluorescein derivatives are detected by a fluorescence spectroscopic method without binding of a reporter group to the tag.

摘要翻译： 提供了包含N4-（取代氨基）胞嘧啶部分的核酸杂交探针，其中取代的氨基包括标签部分，由此检测探针。 还提供了制备本发明的探针的方法，在这些方法中使用的中间体以及在杂交测定中使用本发明的探针的方法。 与本发明的探针一起使用的典型标签部分是通过接头部分连接到探针的修饰胞嘧啶的N4-氨基的生物素，氨基噻二唑和荧光素衍生物。 用生物素标记的探针通常通过结合生物素部分，通过链霉抗生物素蛋白或抗生物素蛋白分子，包括链霉抗生物素蛋白或抗生物素蛋白的报道基团检测，然后检测由于报告基团引起的信号。 用氨基噻二唑衍生物标记的探针通常用与生物素标记的那些基本上相同的方法来检测，但采用作为通过碳酸酐酶分子与衍生物结合的报告基团。 用荧光素衍生物标记的探针通过荧光光谱法检测，而不将报道基团与标签结合。

公开(公告)号：US4302204A

公开(公告)日：1981-11-24

申请号：US54200

申请日：1979-07-02

申请人： Geoffrey M. Wahl ,  George R. Stark

发明人： Geoffrey M. Wahl ,  George R. Stark

IPC分类号： C12N15/10 ,  C12Q1/68 ,  C12Q1/6832 ,  C12Q1/6834 ,  G01N31/22 ,  G01N33/16 ,  G01N33/48

CPC分类号： C12Q1/6832 ,  C12N15/1034 ,  C12Q1/6834 ,  Y10S435/805

摘要： Improvements in the transfer and detection of separated nucleic acids, both RNA and DNA, are provided. For analysis of large DNA, the molecular weight segregated fractions of DNA are depurinated and fragmented to provide fractions having less than about 2 kb as single strands. With both RNA and DNA, the nucleic acid fractions are transferred after resolution to a chemically treated substrate and covalently affixed to the substrate. The resulting nucleotides affixed to the substrate are hybridized with labeled nucleotide probes and a volume exclusion agent, particularly a water soluble ionic polymer.

摘要翻译： 提供分离的核酸（RNA和DNA）的转移和检测的改进。 为了分析大的DNA，DNA的分子量分离级分被去除并分离，以提供具有小于约2kb的单链的级分。 使用RNA和DNA，将核酸级分在分辨后转移到化学处理的底物上并共价固定在底物上。 附着于底物的所得核苷酸与标记的核苷酸探针和体积排阻剂，特别是水溶性离子聚合物杂交。

公开(公告)号：US07371577B2

公开(公告)日：2008-05-13

申请号：US11184156

申请日：2005-07-18

申请人： Geoffrey M. Wahl ,  Stephen V. O'Gorman

发明人： Geoffrey M. Wahl ,  Stephen V. O'Gorman

IPC分类号： C12N15/87 ,  C12N5/00 ,  C12N5/02

CPC分类号： C12N15/90 ,  A61K38/00 ,  A61K48/00 ,  C12N9/00 ,  C12N15/52 ,  C12N15/85 ,  C12N15/907 ,  C12N2800/30

摘要： A gene activation/inactivation and site-specific integration system has been developed for mammalian cells. The invention system is based on the recombination of transfected sequences by FLP, a recombinase derived from Saccharomyces. In several cell lines, FLP has been shown to rapidly and precisely recombine copies of its specific target sequence. For example, a chromosomally integrated, silent β-galactosidase reporter gene was activated for expression by FLP-mediated removal of intervening sequences to generate clones of marked cells. Alternatively, the reverse reaction can be used to target transfected DNA to specific chromosomal sites. These results demonstrate that FLP can be used, for example, to mosaically activate or inactivate transgenes for a variety of therapeutic purposes, as well as for analysis of vertebrate development. The FLP recombination system of the present invention can be incorporated in transgenic, non-human mammals to achieve site-specific integration of transgenes, to construct functional genes or to disrupt existing genes.

摘要翻译： 已经为哺乳动物细胞开发了基因激活/失活和位点特异性整合系统。 本发明系统基于通过FLP转染的序列的重组，其是衍生自酵母属的重组酶。 在几种细胞系中，已经显示FLP快速且精确地重组其特异性靶序列的拷贝。 例如，染色体整合的沉默的β-半乳糖苷酶报道基因通过FLP介导的中间序列的去除被激活用于表达以产生标记细胞的克隆。 或者，反向反应可用于将转染的DNA靶向特定的染色体位点。 这些结果表明，FLP可以用于例如用于各种治疗目的的马赛克激活或灭活转基因，以及用于脊椎动物发育的分析。 本发明的FLP重组系统可并入转基因非人哺乳动物中，以实现转基因的位点特异性整合，构建功能基因或破坏现有基因。

公开(公告)号：US5885836A

公开(公告)日：1999-03-23

申请号：US825784

申请日：1997-04-08

申请人： Geoffrey M. Wahl ,  Stephen V. O'Gorman

发明人： Geoffrey M. Wahl ,  Stephen V. O'Gorman

IPC分类号： A01K67/027 ,  A61K38/00 ,  A61K48/00 ,  C12N9/00 ,  C12N15/52 ,  C12N15/85 ,  C12N15/90 ,  C12Q1/68 ,  C12N15/63 ,  C12N15/87

CPC分类号： C12N15/90 ,  C12N15/52 ,  C12N15/85 ,  C12N15/907 ,  C12N9/00 ,  A61K38/00 ,  A61K48/00 ,  C12N2800/30

摘要： A gene activation/inactivation and site-specific integration system has been developed for mammalian cells. The invention system is based on the recombination of transfected sequences by FLP, a recombinase derived from Saccharomyces. In several cell lines, FLP has been shown to rapidly and precisely recombine copies of its specific target sequence. For example, a chromosomally integrated, silent .beta.-galactosidase reporter gene was activated for expression by FLP-mediated removal of intervening sequences to generate clones of marked cells. Alternatively, the reverse reaction can be used to target transfected DNA to specific chromosomal sites. These results demonstrate that FLP can be used, for example, to mosaically activate or inactivate transgenes for a variety of therapeutic purposes, as well as for analysis of vertebriate development.

摘要翻译： 已经为哺乳动物细胞开发了基因激活/失活和位点特异性整合系统。 本发明系统基于通过FLP转染的序列的重组，其是衍生自酵母属的重组酶。 在几种细胞系中，已经显示FLP快速且精确地重组其特异性靶序列的拷贝。 例如，染色体整合的沉默的β-半乳糖苷酶报道基因通过FLP介导的中间序列的去除被激活用于表达以产生标记细胞的克隆。 或者，反向反应可用于将转染的DNA靶向特定的染色体位点。 这些结果表明，FLP可用于例如用于各种治疗目的的马赛克激活或灭活转基因，以及用于分析叶酸发育。

公开(公告)号：US5677177A

公开(公告)日：1997-10-14

申请号：US486409

申请日：1995-06-07

申请人： Geoffrey M. Wahl ,  Stephen V. O'Gorman

发明人： Geoffrey M. Wahl ,  Stephen V. O'Gorman

IPC分类号： A01K67/027 ,  A61K38/00 ,  A61K48/00 ,  C12N9/00 ,  C12N15/52 ,  C12N15/85 ,  C12N15/90 ,  C12Q1/68 ,  C12N5/06 ,  C12N15/09

CPC分类号： C12N15/90 ,  C12N15/52 ,  C12N15/85 ,  C12N15/907 ,  C12N9/00 ,  A61K38/00 ,  A61K48/00 ,  C12N2800/30

摘要： A gene activation/inactivation and site-specific integration system has been developed for mammalian cells. The invention system is based on the recombination of transfected sequences by FLP, a recombinase derived from Saccharomyces. In several cell lines, FLP has been shown to rapidly and precisely recombine copies of its specific target sequence. For example, a chromosomally integrated, silent .beta.-galactosidase reporter gene was activated for expression by FLP-mediated removal of intervening sequences to generate clones of marked cells. Alternatively, the reverse reaction can be used to target transfected DNA to specific chromosomal sites. These results demonstrate that FLP can be used, for example, to mosaically activate or inactivate transgenes for a variety of therapeutic purposes, as well as for analysis of vertebriate development.

摘要翻译： 已经为哺乳动物细胞开发了基因激活/失活和位点特异性整合系统。 本发明系统基于通过FLP转染的序列的重组，其是衍生自酵母属的重组酶。 在几种细胞系中，已经显示FLP快速且精确地重组其特异性靶序列的拷贝。 例如，染色体整合的沉默的β-半乳糖苷酶报道基因通过FLP介导的中间序列的去除被激活用于表达以产生标记细胞的克隆。 或者，反向反应可用于将转染的DNA靶向特定的染色体位点。 这些结果表明，FLP可用于例如用于各种治疗目的的马赛克激活或灭活转基因，以及用于分析叶酸发育。

公开(公告)号：US5130446A

公开(公告)日：1992-07-14

申请号：US323433

申请日：1989-03-14

申请人： Gary F. Musso ,  Soumitra Ghosh ,  Leslie E. Orgel ,  Geoffrey M. Wahl ,  Emil T. Kaiser

发明人： Gary F. Musso ,  Soumitra Ghosh ,  Leslie E. Orgel ,  Geoffrey M. Wahl ,  Emil T. Kaiser

IPC分类号： C07F7/18 ,  C07H21/00 ,  C12Q1/68

CPC分类号： C07H21/00 ,  C07F7/1852 ,  C07F7/1856 ,  C12Q1/6816

摘要： Fluorescent linker moieties are provided which comprise a fluorescent compound such as fluorescein attached to a linker moiety such that a functional group of the linker is available for attachment to an affinity molecule such as a nucleic acid which has an N.sup.4 (substituted amino) cytosine moiety. Probes tagged with fluorescent derivatives such as fluorescein, tetramethyrhodamine or tetraethylrhodamine may be detected by fluorescence spectroscopic methods.

摘要翻译： 提供荧光接头部分，其包含荧光化合物，例如与连接体部分连接的荧光素，使得接头的官能团可用于连接到亲和分子例如具有N4（取代的氨基）胞嘧啶部分的核酸。 用荧光素，四甲嘧啶胺或四乙基罗丹明等荧光衍生物标记的探针可以通过荧光光谱法检测。

公开(公告)号：US20120076762A1

公开(公告)日：2012-03-29

申请号：US13259644

申请日：2010-03-24

申请人： Teruhisa Kawamura ,  Jotaro Suzuki ,  Yunyuan V. Wang ,  Angel Raya ,  Geoffrey M. Wahl ,  Juan Carlos Izpisua Belmonte

发明人： Teruhisa Kawamura ,  Jotaro Suzuki ,  Yunyuan V. Wang ,  Angel Raya ,  Geoffrey M. Wahl ,  Juan Carlos Izpisua Belmonte

IPC分类号： A61K35/12 ,  C12N5/10 ,  A61P43/00 ,  C12N15/85

CPC分类号： C12N5/0696 ,  C12N2501/40 ,  C12N2501/602 ,  C12N2501/603 ,  C12N2501/998 ,  C12N2510/00

摘要： Methods and compositions are provided for, inter alia, the generation of induced pluripotent stem cells. Induced pluripotent stem cells may be generated by reprogramming and inhibition of p53. Further, useful intermediates for the generation of induced pluripotent stem cells are also provided.

摘要翻译： 提供方法和组合物，尤其是诱导多能干细胞的产生。 诱导的多能干细胞可通过重编程和抑制p53产生。 此外，还提供了用于产生诱导多能干细胞的有用中间体。

公开(公告)号：US06312908B1

公开(公告)日：2001-11-06

申请号：US09519931

申请日：2000-03-07

申请人： Geoffrey M. Wahl ,  Noriaki Shimizu

发明人： Geoffrey M. Wahl ,  Noriaki Shimizu

IPC分类号： C12Q168

CPC分类号： C12Q1/6806 ,  C12N15/1003 ,  C12Q1/6886

摘要： The present invention provides a method for the isolation of extrachromosomal amplified nucleic acids that are associated with a cell proliferative disorder. Isolation and further identification of such genes is critical for diagnosis, prognosis, and course of therapy.

摘要翻译： 本发明提供了分离与细胞增殖性病症相关的染色体外扩增核酸的方法。 这些基因的分离和进一步鉴定对于诊断，预后和治疗过程至关重要。

公开(公告)号：US6033849A

公开(公告)日：2000-03-07

申请号：US704391

申请日：1996-08-26

申请人： Geoffrey M. Wahl ,  Noriaki Shimizu

发明人： Geoffrey M. Wahl ,  Noriaki Shimizu

IPC分类号： C12N15/10 ,  C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/6806 ,  C12N15/1003 ,  C12Q1/6886

摘要： The present invention provides a method for the isolation of extrachromosomal amplified nucleic acids that are associated with a cell proliferative disorder. Isolation and further identification of such genes is critical for diagnosis, prognosis, and course of therapy.