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11. 发明授权

US08993256B2 Fusion proteins and polynucleotide constructs to measure protein turn-over 有权
标题翻译：融合蛋白和多核苷酸构建体来测量蛋白质翻转
公开(公告)号：US08993256B2
公开(公告)日：2015-03-31
申请号：US12443411
申请日：2007-09-28
申请人： Michael Z. Lin , Roger Y. Tsien
发明人： Michael Z. Lin , Roger Y. Tsien
IPC分类号： C12Q1/02 , C12N9/00 , C12N9/48 , C12N1/20 , C12N15/00 , C07H21/04 , C12Q1/37 , G01N33/58
CPC分类号： C12Q1/37 , G01N33/582
摘要： The disclosure provides method and compositions for visualizing protein turnover. In particular, the disclosure provides methods and compositions useful for measuring the age of particular proteins or the dynamics of localized protein translation.
摘要翻译：本公开提供了用于可视化蛋白质周转的方法和组合物。特别地，本公开提供了可用于测量特定蛋白质的年龄或局部蛋白质翻译的动力学的方法和组合物。

12. 发明授权

US08759492B2 Engineered red-shifted channelrhodopsin variants 有权
标题翻译：工程红移通道视紫红质变异体
公开(公告)号：US08759492B2
公开(公告)日：2014-06-24
申请号：US13588957
申请日：2012-08-17
申请人： John Yu-luen Lin , Roger Y. Tsien
发明人： John Yu-luen Lin , Roger Y. Tsien
IPC分类号： A61K36/00 , C12P21/08
CPC分类号： C07K19/00 , A61K48/005 , A61N5/062 , C07K14/705 , C07K2319/00 , C12N15/62 , C12N15/85 , C12N15/864 , C12N2740/15043 , C12N2740/15071 , C12N2750/14143 , C12N2750/14171
摘要： The invention provides engineered red-shifted channelrhodopsin variants. In some embodiments, the channelrhodopsin variants are characterized by improved membrane trafficking, expression, and/or unique spectral and kinetic properties.
摘要翻译：本发明提供工程化的红移通道视紫红质变体。在一些实施方案中，通道视紫红质变体的特征在于改进的膜运输，表达和/或独特的光谱和动力学性质。

13. 发明申请

US20140093907A1 FLUORESCENT PROBES FOR MONITORING VOLTAGE BY PHOTO-INDUCED ELECTRON TRANSFER 有权
标题翻译：用于通过光电导体传输监测电压的荧光探针
公开(公告)号：US20140093907A1
公开(公告)日：2014-04-03
申请号：US14118836
申请日：2012-05-21
申请人： Evan Walker Miller , Roger Y. Tsien
发明人： Evan Walker Miller , Roger Y. Tsien
IPC分类号： G01N21/64
CPC分类号： G01N21/6486 , C09K11/06 , C09K2211/1007 , C09K2211/1011 , C09K2211/1088 , G01N33/5008 , G01N33/542 , G01N2500/10
摘要： Compounds and methods for determining transmembrane potential, monitoring changes in transmembrane potential, and/or drug screening are provided. In one aspect, compounds of the invention have a structure according to the formula: E-M-A, wherein A is a fluorophore, selected from xanthenes, eoumarins, cyanines, bimanes, and difluoroboradizaindacenes, charged at physiological pH; M is a molecular wire; and E is a hydrophobic moiety, wherein A and E are capable of being involved in a photo-induced, intramolecular electron transfer that quenches the fluorescence of A in response to a voltage condition. When in use, compounds of the invention are membrane-impermeant and oriented within the cell membrane such that the charged moiety localizes at the outer leaflet of the lipid bilayer and the hydrophobic moiety and molecular wire associate with the hydrophobic portion of the lipid bilayer. The rate of electron transfer, fluorescence intensity, and quenching are altered in response to changes in transmembrane potential.
摘要翻译：提供了确定跨膜电位的化合物和方法，监测跨膜电位的变化和/或药物筛选。在一个方面，本发明的化合物具有下式的结构：其中A是荧光团，选自在生理pH下加入的呫吨，香豆素，花青，双马来酰亚胺和二氟硼烷二恶烷; M是分子线; 并且E是疏水部分，其中A和E能够参与光诱导的分子内电子转移，其响应于电压条件淬灭A的荧光。当使用时，本发明的化合物是膜不渗透的并且在细胞膜内定向，使得带电部分定位在脂质双层的外部小叶上，并且疏水部分和分子线与脂质双层的疏水部分缔合。响应于跨膜电位的变化，电子转移速率，荧光强度和淬灭速率发生变化。

14. 发明申请

US20110076711A1 PHOTON REDUCING AGENTS FOR FLUORESCENCE ASSAYS 有权
标题翻译：用于荧光测定的光子还原剂
公开(公告)号：US20110076711A1
公开(公告)日：2011-03-31
申请号：US12861791
申请日：2010-08-23
申请人： Tom KNAPP , Gregor Zlokarnik , Paul Negulescu , Roger Y. Tsien , Timothy James Rink
发明人： Tom KNAPP , Gregor Zlokarnik , Paul Negulescu , Roger Y. Tsien , Timothy James Rink
IPC分类号： C12Q1/02
CPC分类号： G01N33/582 , C12Q1/682 , G01N33/5005 , G01N33/542 , Y10S436/80 , Y10S436/809 , Y10T436/13 , C12Q2563/107 , C12Q2527/125
摘要： The present invention provides a method for reducing undesirable light emission from a sample using at least one photon producing agent and at least one photon reducing agent (e.g. dye-based photon reducing agents). The present invention further provides a method for reducing undesirable light emission from a sample (e.g., a biochemical or cellular sample) with at least one photon producing agent and at least one collisional quencher. The present invention also provides a method for reducing undesirable light emission from a sample (e.g., a biochemical or cellular sample) with at least one photon producing agent and at least one quencher, such as an electronic quencher. The present invention also provides a system and method of screening test chemicals in fluorescent assays using photon reducing agents. The present invention also provides compositions, pharmaceutical compositions, and kits for practicing these methods.
摘要翻译：本发明提供一种使用至少一种光子产生剂和至少一种光子还原剂（例如基于染料的光子还原剂）来减少来自样品的不希望的光发射的方法。本发明进一步提供了用至少一种光子产生剂和至少一种碰撞猝灭剂减少来自样品（例如生物化学或细胞样品）的不希望的光发射的方法。本发明还提供了用至少一种光子产生剂和至少一种猝灭剂（例如电子猝灭剂）减少来自样品（例如，生物化学或细胞样品）的不希望的光发射的方法。本发明还提供了使用光子还原剂在荧光测定中筛选测试化学品的系统和方法。本发明还提供了用于实施这些方法的组合物，药物组合物和试剂盒。

15. 发明授权

US07854898B2 Multi-well platforms, caddies, lids and combinations thereof 有权
标题翻译：多井平台，球场，盖子及其组合
公开(公告)号：US07854898B2
公开(公告)日：2010-12-21
申请号：US12271762
申请日：2008-11-14
申请人： Andrew A. Pham , Peter J. Coassin , Alec Tate Harootunian , Peter N. Pham , Harry Stylli , Roger Y. Tsien
发明人： Andrew A. Pham , Peter J. Coassin , Alec Tate Harootunian , Peter N. Pham , Harry Stylli , Roger Y. Tsien
IPC分类号： B01L3/00
CPC分类号： B01L3/5085 , B01J2219/00315 , B01J2219/00317 , B01J2219/00659 , B01J2219/00707 , B01J2219/0072 , B01L3/50855 , B01L9/523 , B01L2300/0654 , B01L2300/0829 , C12M23/12 , C40B60/14
摘要： The present invention relates to multi-well platforms, lids, caddies and any combination thereof. The multi-well platforms comprise a plurality of wells that can be of any shape and can be arranged in any ornamental pattern. The lid comprises an area corresponding to the well field. The caddy has a footprint approximately that of a standard 96-well microtiter plate. When the multi-well platform is engaged with the caddy, the wells of the well-field of the multi-well platform are preferably not obscured by the caddy when viewed from the bottom of the combination.
摘要翻译：本发明涉及多孔平台，盖子，球童及其任何组合。多井平台包括可以是任何形状的多个井，并且可以以任何装饰图案排列。盖子包括对应于井场的区域。球罐的尺寸大约是标准96孔微孔板的面积。当多孔平台与球童接合时，当从组合的底部观察时，多孔平台的井场的井优选地不被球童遮蔽。

16. 发明申请

US20080293068A1 METHODS FOR ENGINEERING POLYPEPTIDE VARIANTS VIA SOMATIC HYPERMUTATION AND POLYPEPTIDE MADE THEREBY 有权
标题翻译：通过超声波和多肽制备多肽变体的方法
公开(公告)号：US20080293068A1
公开(公告)日：2008-11-27
申请号：US12124888
申请日：2008-05-21
申请人： Roger Y. Tsien , Lei Wang
发明人： Roger Y. Tsien , Lei Wang
IPC分类号： C12Q1/68 , C12P21/04 , C12Q1/02
CPC分类号： C12P21/02
摘要： Methods using somatic hypermutation (SHM) for producing polypeptide and nucleic acid variants, and nucleic acids encoding such polypeptide variants are disclosed. Such variants may have desired properties. Also disclosed are novel polypeptides, such as improved fluorescent proteins, produced by the novel methods, and nucleic acids, vectors, and host cells comprising such vectors.
摘要翻译：公开了使用体细胞超突变（SHM）用于产生多肽和核酸变体的方法，以及编码这种多肽变体的核酸。这样的变体可以具有期望的性质。还公开了通过新方法产生的新型多肽，例如改良的荧光蛋白，以及包含这种载体的核酸，载体和宿主细胞。

17. 发明授权

US07173130B2 Detection of transmembrane potentials by optical methods 有权
标题翻译：通过光学方法检测跨膜电位
公开(公告)号：US07173130B2
公开(公告)日：2007-02-06
申请号：US09967772
申请日：2001-09-28
申请人： Roger Y. Tsien , Jesus E. Gonzalez, III
发明人： Roger Y. Tsien , Jesus E. Gonzalez, III
IPC分类号： C07D403/06
CPC分类号： G01N33/502 , C07D209/08 , C07D215/227 , C07D239/60 , C07D239/62 , C07D239/66 , C07D239/80 , C07D241/44 , C07D265/36 , C07D279/16 , C07D311/16 , C07D487/04 , C07F5/003 , C07F5/02 , C07F5/027 , C07F9/10 , C07F9/5728 , C07K2319/033 , C09B23/02 , G01N33/5005 , G01N33/5008 , G01N33/5044 , G01N33/5058 , G01N33/5061 , G01N33/5076 , G01N33/52 , G01N33/542 , G01N33/582 , G01N33/6872 , G01N33/92 , G01N2405/04 , G01N2500/10 , Y10S436/80 , Y10S436/805
摘要： Methods and compositions are provided for detecting changes in membrane potential in membranes biological systems. In one aspect, the method comprises; a) providing a living cell with a first reagent comprising a charged hydrophobic molecule which is typically a fluorescence resonance energy transfer (FRET) acceptor or donor, or is a quencher and is capable of redistributing within the membrane of a biological membrane in response to changes in the potential across the membrane; b) providing the cell with a second reagent that can label the first face or the second face of a biological membrane within the cell; c) detecting light emission from the first reagent or the second reagent. One aspect of this method involves monitoring membrane potential changes in subcellular organelle membranes in a living cells.Another aspect of the invention is the use of certain embodiments of the method for the screening of test chemicals for activity to modulate the activity of a target ion channel.Another aspect of the present invention is a transgenic organism comprising a first reagent that comprises a charged hydrophobic fluorescent molecule, and a second reagent comprising a bioluminescent or naturally fluorescent protein.
摘要翻译：提供了用于检测膜生物系统中膜电位变化的方法和组合物。一方面，该方法包括： a）向活细胞提供包含通常为荧光共振能量转移（FRET）受体或供体的带电荷的疏水性分子的第一试剂，或者是猝灭剂，并且能够响应于变化而重新分布在生物膜的膜内在膜的潜力; b）向细胞提供可以标记细胞内生物膜的第一面或第二面的第二试剂; c）检测来自第一试剂或第二试剂的发光。该方法的一个方面涉及监测活细胞中亚细胞细胞器膜中的膜电位变化。本发明的另一方面是该方法的某些实施方案用于筛选测试化学品以调节靶离子通道的活性的活性。本发明的另一方面是转基因生物，其包含含有带电荷的疏水荧光分子的第一试剂和包含生物发光或天然荧光蛋白的第二试剂。

18. 发明授权

US07138503B2 Synthetic molecules that specifically react with target sequences 有权
公开(公告)号：US07138503B2
公开(公告)日：2006-11-21
申请号：US10772164
申请日：2004-02-03
申请人： Roger Y. Tsien , B. Albert Griffin
发明人： Roger Y. Tsien , B. Albert Griffin
IPC分类号： C07K16/00 , C07K7/06 , C07K7/08
CPC分类号： C07F9/74 , C07F9/80 , G01N33/532
摘要： The present invention features biarsenical molecules. Target sequences that specifically react with the biarsenical molecules are also included. The present invention also features kits that include biarsenical molecules and target sequences. Tetraarsenical molecules are also featured in the invention.

19. 发明授权

US06852849B2 Non-oligomerizing tandem fluorescent proteins 有权
标题翻译：非低聚串联荧光蛋白
公开(公告)号：US06852849B2
公开(公告)日：2005-02-08
申请号：US09866538
申请日：2001-05-24
申请人： Roger Y. Tsien , Robert E. Campbell
发明人： Roger Y. Tsien , Robert E. Campbell
IPC分类号： C07K14/435 , C12Q1/25 , G01N33/58 , G01N33/84 , C07H21/04 , C07K14/00 , C12N15/09
CPC分类号： G01N33/587 , C07K14/43595 , C12Q1/25 , G01N33/84 , G01N2500/02
摘要： Non-oligomerizing fluorescent proteins, which are formed by operatively linking two or more monomers of a fluorescent protein, or which are derived from a fluorescent protein having at least one mutation that reduces or eliminates the ability of the fluorescent protein to oligomerize, are provided. The non-oligomerizing fluorescent proteins can be derived from a naturally occurring green fluorescent protein, a red fluorescent protein, or other fluorescent protein, or a fluorescent protein related thereto. Also provided is a fusion protein, which includes a non-oligomerizing fluorescent protein linked to at least one polypeptide of interest. In addition, a polynucleotide encoding a non-oligomerizing fluorescent protein is provided, as is a recombinant nucleic acid molecule, which includes polynucleotide encoding a non-oligomerizing fluorescent protein operatively linked to at least a second polynucleotide. Vectors and host cells containing such polynucleotides also are provided, as are kits containing one or more non-oligomerizing fluorescent proteins or encoding polynucleotides or constructs derived therefrom. Further provided are methods of making and using the proteins and polynucleotides.
摘要翻译：提供了通过可操作地连接两种或更多种荧光蛋白单体或衍生自具有降低或消除荧光蛋白低聚的能力的至少一种突变的荧光蛋白而形成的非低聚荧光蛋白。非低聚荧光蛋白可以衍生自天然存在的绿色荧光蛋白，红色荧光蛋白或其他荧光蛋白，或与其相关的荧光蛋白。还提供了融合蛋白，其包括与至少一种目的多肽连接的非低聚荧光蛋白。此外，提供编码非低聚荧光蛋白的多核苷酸，重组核酸分子也包括编码与至少第二个多核苷酸有效连接的非低聚荧光蛋白的多核苷酸。还提供了含有这种多核苷酸的载体和宿主细胞，以及含有一种或多种非低聚荧光蛋白或编码多核苷酸或由其衍生的构建体的试剂盒。进一步提供制备和使用蛋白质和多核苷酸的方法。

20. 发明授权

US06730520B2 Low fluorescence assay platforms and related methods for drug discovery 有权
标题翻译：低荧光测定平台及药物发现的相关方法
公开(公告)号：US06730520B2
公开(公告)日：2004-05-04
申请号：US10120644
申请日：2002-04-09
申请人： Peter J. Coassin , Alec Tate Harootunian , Andrew A. Pham , Harry Stylli , Roger Y. Tsien
发明人： Peter J. Coassin , Alec Tate Harootunian , Andrew A. Pham , Harry Stylli , Roger Y. Tsien
IPC分类号： G01N2176
CPC分类号： B01L3/5085 , B01J2219/00315 , B01J2219/00317 , B01J2219/00659 , B01J2219/00707 , B01J2219/0072 , B01L9/523 , B01L2300/021 , B01L2300/0829 , B01L2300/12 , C12M23/12 , C12Q1/00 , C40B60/14 , G01N1/30 , G01N21/03 , G01N21/6428 , G01N21/6452 , G01N33/5308 , G01N33/542 , G01N2021/0346
摘要： One aspect of the present invention is a multi-well platform for fluorescence measurements, comprising a plurality of wells within a frame, wherein the multi-well platform has low fluorescence background. Another aspect of the present invention is a system for spectroscopic measurements, comprising reagents for an assay and a multi-well platform for fluorescence measurements. A further aspect of the present invention is a method for detecting the presence of an analyte in a sample contained in a multi-well platform by detecting light emitted from the sample. Another aspect of the present invention is a method from identifying a modulator of a biological process or target in a sample contained in a multi-well platform by detecting light emitted from the sample. Another aspect of the present invention is a composition identified by this method. A further aspect of the present invention is a method to identify a therapeutic. A further aspect of the present invention is a method of testing a therapeutic for therapeutic activity and toxicology by identifying a therapeutic using a method of the present invention and monitoring the toxicology and efficacy of the therapeutic in an in vivo model.
摘要翻译：本发明的一个方面是用于荧光测量的多孔平台，其包括框架内的多个孔，其中所述多孔平台具有低荧光背景。本发明的另一方面是用于光谱测量的系统，其包括用于测定的试剂和用于荧光测量的多孔平台。本发明的另一方面是通过检测从样品发射的光来检测多孔平台中包含的样品中分析物的存在的方法。本发明的另一方面是通过检测从样品发射的光来识别多孔平台中包含的样品中生物过程或靶的调节剂的方法。本发明的另一方面是通过该方法鉴定的组合物。本发明的另一方面是鉴定治疗剂的方法。本发明的另一方面是通过使用本发明的方法鉴定治疗剂并监测治疗剂在体内模型中的毒理学和功效来测试治疗活性和毒理学治疗剂的方法。

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