WO2001036617A3 - Assay for measuring enzyme activity in vivo - Google Patents

Assay for measuring enzyme activity in vivo Download PDF

Info

Publication number
WO2001036617A3
WO2001036617A3 PCT/GB2000/004348 GB0004348W WO0136617A3 WO 2001036617 A3 WO2001036617 A3 WO 2001036617A3 GB 0004348 W GB0004348 W GB 0004348W WO 0136617 A3 WO0136617 A3 WO 0136617A3
Authority
WO
WIPO (PCT)
Prior art keywords
binding domain
binding partner
binding
vivo
organism
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/GB2000/004348
Other languages
French (fr)
Other versions
WO2001036617A2 (en
Inventor
Roger Kingdon Craig
Simon Green
John Colyer
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Cyclacel Ltd
Celltech R&D Ltd
Original Assignee
Cyclacel Ltd
Celltech R&D Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Cyclacel Ltd, Celltech R&D Ltd filed Critical Cyclacel Ltd
Priority to AU14022/01A priority Critical patent/AU1402201A/en
Priority to EP00976136A priority patent/EP1232255A2/en
Publication of WO2001036617A2 publication Critical patent/WO2001036617A2/en
Publication of WO2001036617A3 publication Critical patent/WO2001036617A3/en
Priority to US10/147,354 priority patent/US6852906B2/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0045—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent agent being a peptide or protein used for imaging or diagnosis in vivo
    • A61K49/0047—Green fluorescent protein [GFP]
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/027—New or modified breeds of vertebrates
    • A01K67/0275—Genetically modified vertebrates, e.g. transgenic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
    • A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0045—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent agent being a peptide or protein used for imaging or diagnosis in vivo
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2217/00—Genetically modified animals
    • A01K2217/05—Animals comprising random inserted nucleic acids (transgenic)
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2227/00—Animals characterised by species
    • A01K2227/10—Mammal
    • A01K2227/105—Murine
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00—Animals characterised by purpose
    • A01K2267/02—Animal zootechnically ameliorated
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00—Animals characterised by purpose
    • A01K2267/03—Animal model, e.g. for test or diseases
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00—Animals characterised by purpose
    • A01K2267/03—Animal model, e.g. for test or diseases
    • A01K2267/0393—Animal model comprising a reporter system for screening tests

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Zoology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Biotechnology (AREA)
  • Pathology (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • General Engineering & Computer Science (AREA)
  • Environmental Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Radiology & Medical Imaging (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Physics & Mathematics (AREA)
  • Animal Husbandry (AREA)
  • Endocrinology (AREA)
  • Urology & Nephrology (AREA)
  • Biophysics (AREA)
  • Diabetes (AREA)
  • Plant Pathology (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Microbiology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Rheumatology (AREA)
  • Biochemistry (AREA)
  • Toxicology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

A method is provided for measuring in vivo in a transgenic non-human multicellular organism the activity of a cellular enzyme, which organism is transgenic by virtue of comprising one or more nucleic acid constructs encoding a binding domain and a binding partner thereof, wherein: (i) the binding domain and/or binding partner comprise a site subject to post-translational modification by the cellular enzyme; (ii) modification of the site by the enzyme affects the interaction between the binding domain and binding partner; and (iii) the binding domain and binding partner each comprise a detectable label such that when the binding domain and binding partner interact, a detectable physical characteristic of one or both of the labels is altered, which method comprises measuring the interaction between the binding domain and the binding partner by measuring changes in said physical characteristic in one or more cells of the transgenic organism. A transgenic non-human multicellular organism is also provided.
PCT/GB2000/004348 1999-11-18 2000-11-15 Assay for measuring enzyme activity in vivo Ceased WO2001036617A2 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU14022/01A AU1402201A (en) 1999-11-18 2000-11-15 Assay for measuring enzyme activity in vivo
EP00976136A EP1232255A2 (en) 1999-11-18 2000-11-15 Assay for measuring enzyme activity in vivo
US10/147,354 US6852906B2 (en) 1999-11-18 2002-05-16 Assay for measuring enzyme activity in vivo

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB9927331.0A GB9927331D0 (en) 1999-11-18 1999-11-18 Assay for measuring enzyme activity in vivo
GB9927331.0 1999-11-18

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US10/147,354 Continuation-In-Part US6852906B2 (en) 1999-11-18 2002-05-16 Assay for measuring enzyme activity in vivo

Publications (2)

Publication Number Publication Date
WO2001036617A2 WO2001036617A2 (en) 2001-05-25
WO2001036617A3 true WO2001036617A3 (en) 2002-05-02

Family

ID=10864769

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2000/004348 Ceased WO2001036617A2 (en) 1999-11-18 2000-11-15 Assay for measuring enzyme activity in vivo

Country Status (5)

Country Link
US (1) US6852906B2 (en)
EP (1) EP1232255A2 (en)
AU (1) AU1402201A (en)
GB (1) GB9927331D0 (en)
WO (1) WO2001036617A2 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7619059B2 (en) 2003-07-29 2009-11-17 Life Technologies Corporation Bimolecular optical probes

Families Citing this family (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1290215A2 (en) * 2000-06-07 2003-03-12 Cyclacel Limited Methods for monitoring enzyme activity
EP1317564B1 (en) * 2000-09-14 2007-08-01 Mount Sinai School of Medicine Screening methods to identify g-proteins and other compounds which modulate phosphodiesterase (pde) activity
CA2445420A1 (en) 2003-07-29 2005-01-29 Invitrogen Corporation Kinase and phosphatase assays
EP1869206A4 (en) * 2005-03-02 2011-08-10 Einstein Coll Med ENZYMATIC SENSORS COMPRISING ENVIRONMENTALLY OR FLUORESCENTLY SENSITIVE LABELS AND USES THEREOF
JP2009513681A (en) * 2005-10-28 2009-04-02 インヴィトロジェン コーポレーション Kinase and ubiquitination assays
WO2008065540A2 (en) 2006-11-27 2008-06-05 Cis Bio International Method for the detection of post-translational modifications
WO2008109161A2 (en) * 2007-03-07 2008-09-12 The Albert Einstein College Of Medicine Of Yeshiva University Deeply quenched enzyme sensors
US20100081926A1 (en) * 2008-09-29 2010-04-01 Searete Llc, A Limited Liability Corporation Of The State Of Delaware Histological facilitation systems and methods
US20100081916A1 (en) * 2008-09-29 2010-04-01 Searete Llc, A Limited Liability Corporation Of The State Of Delaware. Histological facilitation systems and methods
US20100081915A1 (en) * 2008-09-29 2010-04-01 Searete Llc, Alimited Liability Corporation Of The State Of Delaware Histological facilitation systems and methods
US20100081924A1 (en) * 2008-09-29 2010-04-01 Searete Llc, A Limited Liability Corporation Of The State Of Delaware Histological facilitation systems and methods
US20100081927A1 (en) * 2008-09-29 2010-04-01 Searete Llc, A Limited Liability Corporation Of The State Of Delaware Histological facilitation systems and methods
US20100081928A1 (en) * 2008-09-29 2010-04-01 Searete Llc, A Limited Liability Corporation Of The State Of Delaware Histological Facilitation systems and methods
WO2010093916A1 (en) * 2009-02-14 2010-08-19 Genentech, Inc. Methods for the detection of fatty-acylated proteins
CN103930165B (en) 2011-09-02 2018-05-25 纽约市哥伦比亚大学理事会 CaMKII, IP3R, calcineurin, P38 and MK2/3 inhibitors for the treatment of metabolic disorders in obesity
US20140099654A1 (en) 2012-09-26 2014-04-10 Promega Corporation Real-time monitoring

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997028261A1 (en) * 1996-01-31 1997-08-07 The Regents Of The University Of California Tandem fluorescent protein constructs
WO1998030715A1 (en) * 1997-01-07 1998-07-16 California Institute Of Technology Optical sensors of cell signaling
WO1998036081A2 (en) * 1997-02-13 1998-08-20 Memorial Sloan-Kettering Cancer Center Hybrid molecules for optically detecting changes in cellular microenvironments
WO1999018856A1 (en) * 1997-10-10 1999-04-22 Cytovia, Inc. Novel fluorescent reporter molecules and their applications including assays for caspases
EP0949269A1 (en) * 1998-04-02 1999-10-13 SymBiosis GmbH Biosensor protein

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5786158A (en) 1992-04-30 1998-07-28 Yale University Therapeutic and diagnostic methods and compositions based on notch proteins and nucleic acids
WO1997027876A1 (en) 1996-02-02 1997-08-07 Sidney Kimmel Cancer Center Immunotherapy sensitization of cancer therapy
AU6952498A (en) 1997-04-04 1998-10-30 Millenium Pharmaceuticals, Inc. Novel human delta3 compositions and therapeutic and diagnostic uses therefor

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997028261A1 (en) * 1996-01-31 1997-08-07 The Regents Of The University Of California Tandem fluorescent protein constructs
WO1998030715A1 (en) * 1997-01-07 1998-07-16 California Institute Of Technology Optical sensors of cell signaling
WO1998036081A2 (en) * 1997-02-13 1998-08-20 Memorial Sloan-Kettering Cancer Center Hybrid molecules for optically detecting changes in cellular microenvironments
WO1999018856A1 (en) * 1997-10-10 1999-04-22 Cytovia, Inc. Novel fluorescent reporter molecules and their applications including assays for caspases
EP0949269A1 (en) * 1998-04-02 1999-10-13 SymBiosis GmbH Biosensor protein

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
HAN HTUN ET AL: "VISUALIZATION OF GLUCOCORTICOID RECEPTOR TRANSLOCATION AND INTRANUCLEAR ORGANIZATION IN LIVING CELLS WITH A GREEN FLUORESCENT PROTEIN CHIMERA", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA, NATIONAL ACADEMY OF SCIENCE. WASHINGTON, US, vol. 93, no. 10, 1 May 1996 (1996-05-01), pages 4845 - 4850, XP002029560, ISSN: 0027-8424 *
HEIM R ET AL: "ENGINEERING GREEN FLUORESCENT PROTEIN FOR IMPROVED BRIGHTNESS, LONGER WAVELENGTHS AND FLUROESCENCE RESONANCE ENERGY TRANSFER", CURRENT BIOLOGY, CURRENT SCIENCE,, GB, vol. 6, no. 2, 1 February 1996 (1996-02-01), pages 178 - 182, XP000676582, ISSN: 0960-9822 *
MITRA R D ET AL: "FLUORESCENCE RESONANCE ENERGY TRANSFER BETWEEN BLUE-EMITTING AND RED-SHIFTED EXCITATION DERIVATIVES OF THE GREEN FLUORESCENT PROTEIN", GENE: AN INTERNATIONAL JOURNAL ON GENES AND GENOMES, ELSEVIER SCIENCE PUBLISHERS, BARKING, GB, vol. 173, no. 1, 1 July 1996 (1996-07-01), pages 13 - 17, XP002033688, ISSN: 0378-1119 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7619059B2 (en) 2003-07-29 2009-11-17 Life Technologies Corporation Bimolecular optical probes

Also Published As

Publication number Publication date
WO2001036617A2 (en) 2001-05-25
EP1232255A2 (en) 2002-08-21
US20030079235A1 (en) 2003-04-24
US6852906B2 (en) 2005-02-08
GB9927331D0 (en) 2000-01-12
AU1402201A (en) 2001-05-30

Similar Documents

Publication Publication Date Title
WO2001036617A3 (en) Assay for measuring enzyme activity in vivo
ID28595A (en) MODIFIED CARRIER, CATALYSOXIDE COMPLEX AND ACRYLIC ACID MAKING PROCESS
WO2001079463A3 (en) Nucleic acids encoding polypeptides having haloperoxidase activity
WO2004007692A3 (en) Methods and compositions for analyzing polymers using chimeric tags
AU6042699A (en) Assays for measuring nucleic acid binding proteins and enzyme activities
WO2000060060A3 (en) Polypeptides having alkaline alpha-amylase activity and nucleic acids encoding same
WO2000060058A3 (en) Polypeptides having alkaline alpha-amylase activity and nucleic acids encoding same
WO2002016583A3 (en) Constructs and their use in metabolic pathway engineering
AU3246300A (en) Polypeptides having galactose oxidase activity and nucleic acids encoding same
AU2001271919A1 (en) Detection of binding reactions using catalytic electrochemical labels
WO2002004488A3 (en) Nucleic acid binding polypeptides
AUPR975201A0 (en) Enzymatic redox labelling of nucleic acids
AU2001241674A1 (en) Variant galactose oxidase, nucleic acid encoding same, and methods of using same
WO2002102825A3 (en) Fluorescently labelled locked nucleic acids
ATE304595T1 (en) LIGHT PRODUCING BIOSENSOR
WO2003093409A3 (en) Promoters exhibiting endothelial cell specificity and methods of using same
WO2003085082A3 (en) Nucleic acid reactions using labels with different redox potentials
WO2003093429A3 (en) Nucleic acid reactions using labels with different redox potentials
WO2000028043A3 (en) Polypeptides having lactonohydrolase activity and nucleic acids encoding same
AU2002243235A1 (en) Novel nucleic acids, methods and transformed cells for the modulation of gibberellin production
WO2002040629A3 (en) Promoters exhibiting endothelial cell specificity and methods of using same
ATE412056T1 (en) NUCLEIC ACID MOLECULES ENCODING ENZYMES POSSESSING FRUCTOSYL TRANSFERASE ACTIVITY AND THEIR USE
AU2001275389A1 (en) Nucleic acid sequences encoding type iii tenebrio antifreeze proteins and method for assaying activity
WO2000046375A3 (en) Methods for producing polypeptides in fungal cells
AU2813200A (en) Assay for measuring different enzyme activities simultaneously

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 10147354

Country of ref document: US

WWE Wipo information: entry into national phase

Ref document number: 2000976136

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 2000976136

Country of ref document: EP

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWW Wipo information: withdrawn in national office

Ref document number: 2000976136

Country of ref document: EP

DPE2 Request for preliminary examination filed before expiration of 19th month from priority date (pct application filed from 20040101)