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WO2002036784A1 - Animaux transgeniques utilises pour analyser la regulation du gene p450 du cytochrome cyp3a4 - Google Patents

Animaux transgeniques utilises pour analyser la regulation du gene p450 du cytochrome cyp3a4 Download PDF

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Publication number
WO2002036784A1
WO2002036784A1 PCT/AU2001/001407 AU0101407W WO0236784A1 WO 2002036784 A1 WO2002036784 A1 WO 2002036784A1 AU 0101407 W AU0101407 W AU 0101407W WO 0236784 A1 WO0236784 A1 WO 0236784A1
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Prior art keywords
nucleic acid
acid molecule
reporter
human
transcription
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Inventor
Christopher Liddle
Bryan James Goodwin
Graham Robertson
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University of Sydney
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University of Sydney
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Priority claimed from AUPR1161A external-priority patent/AUPR116100A0/en
Priority claimed from AUPR4901A external-priority patent/AUPR490101A0/en
Application filed by University of Sydney filed Critical University of Sydney
Priority to CA2426959A priority Critical patent/CA2426959C/fr
Priority to AU2002213660A priority patent/AU2002213660A1/en
Priority to US10/415,607 priority patent/US7531712B2/en
Priority to JP2002539530A priority patent/JP2004512054A/ja
Priority to EP01981959A priority patent/EP1337650A4/fr
Publication of WO2002036784A1 publication Critical patent/WO2002036784A1/fr
Anticipated expiration legal-status Critical
Priority to US12/430,672 priority patent/US20090307785A1/en
Priority to US12/463,246 priority patent/US8088968B2/en
Priority to US12/855,383 priority patent/US8629318B2/en
Ceased legal-status Critical Current

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    • 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
    • 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
    • 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
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • C12N9/0077—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14) with a reduced iron-sulfur protein as one donor (1.14.15)
    • 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
    • A01K2267/03—Animal model, e.g. for test or diseases
    • A01K2267/0393—Animal model comprising a reporter system for screening tests

Definitions

  • Such an animal model would not be useful unless at least some of the aspects of the regulation of CYP3A4 gene expression in the human, especially tissue specific expression, are reproduced. This is because in the human, the CYP3A4 gene is expressed in specific tissues, including liver and small intestine, which many compounds inevitably come into contact with when administered for the purpose of therapy. Accordingly, one would be unable to determine whether the bio-availability of a candidate drug would be sufficient for achieving an intended therapeutic effect in clinical practice in a model which does not reproduce the constitutive and xenobiotic induced tissue specific expression of the CYP3A4 gene that is observed in the human.
  • the invention seeks to address the above identified need and in a first aspect provides a non-human mammal comprising:
  • a reporter nucleic acid molecule for producing a detectable amount of a reporter molecule for indicating regulation of transcription of the reporter nucleic acid molecule by the regulatory nucleic acid molecule
  • the inventors have found that the incorporation of a region of the human CYP3A4 gene that is located between the initiation of transcription site of the gene and a position 13,000 nucleotides upstream of the initiation of transcription site into an animal model provides the animal with sufficient genetic information for reproducing the constitutive and xenobiotic induced tissue specific expression of the CYP3A4 gene that is observed in humans. More specifically, the inventors have generated animal models which contain a transgene comprising this region and have observed that these models provide constitutive and xenobiotic inducible expression of a transgene in a tissue pattern which reproduces the tissue specific expression of CYP3A4 which is observed in a human. Importantly, the level of constitutive expression is sufficient to allow one to observe the effect on the regulation of tissue specific transgene expression, of administration of a compound, for example, a xenobiotic or steroid, to the animal.
  • the inventors have observed that the animal models described herein also reproduce aspects of the constitutive and xenobiotic inducible developmental expression of the CYP3A4 gene that is observed in humans.
  • the invention provides a non human mammal comprising:
  • a regulatory nucleic acid molecule comprising a nucleotide sequence that is identical to the nucleotide sequence of the human CYP3A4 gene that extends about 13,000 nucleotides upstream from the initiation of transcription site of the gene;
  • a reporter nucleic acid molecule for producing a detectable amount of a reporter molecule for indicating regulation of transcription of the reporter nucleic acid molecule by the regulatory nucleic acid molecule
  • reporter and regulatory nucleic acid molecules are arranged to permit the regulatory nucleic acid molecule to regulate transcription of the reporter nucleic acid molecule.
  • the regulatory nucleic acid molecule comprises the sequence shown in SEQ ID NO : 1.
  • the inventors have generated transgenic animals which contain a region of the human CYP3A4 gene between the initiation of transcription site and a position about 3,200 nucleotides upstream of the initiation transcription site and observed that the transgene is not constitutively expressed or inducible by xenobiotics in these animals. Accordingly, the inventors have found that the genetic information required for reproducing the constitutive and xenobiotic induced tissue specific and developmental expression of CYP3A4 observed in a human is contained in the region of the human CYP3A4 gene between the position located about 3,200 nucleotides upstream of the initiation of transcription site of the gene and a position 13,000 nucleotides upstream of the initiation of transcription site.
  • the invention provides a non-human mammal comprising:
  • a regulatory nucleic acid molecule comprising a nucleotide sequence that is identical to the sequence of the human CYP3A4 gene that extends about 8,000 nucleotides upstream from a position about 3,000 nucleotides upstream from the initiation of transcription site of the gene;
  • reporter and regulatory nucleic acid molecules are arranged to permit the regulatory nucleic acid molecule to regulate transcription of the reporter nucleic acid molecule.
  • the regulatory nucleic acid molecule comprises the sequence shown in SEQ ID NO:2.
  • the invention provides a non-human mammal comprising:
  • a regulatory nucleic acid molecule which is capable of regulating transcription of the human CYP3A4 gene and which comprises a nucleotide sequence that is identical to the sequence of the human CYP3A4 gene that extends about 600 nucleotides upstream from a position about 7,200 nucleotides upstream of the initiation of transcription site of the gene;
  • a reporter nucleic acid molecule for producing a detectable amount of a reporter molecule for indicating regulation of transcription of the reporter nucleic acid molecule by the regulatory nucleic acid molecule
  • reporter and regulatory nucleic acid molecules are arranged to permit the regulatory nucleic acid molecule to regulate transcription of the reporter nucleic acid molecule.
  • the regulatory nucleic acid molecule comprises the sequence shown in SEQ ID NO: 3.
  • the regulatory nucleic acid molecule has the sequence of any one of the following fragments of the CYP3A4 gene:
  • a regulatory nucleic acid molecule which has the sequence of a fragment consisting of from nucleotide positions -7836 to -7207 contiguous with -362 to +53 is particularly preferred, as this construct contains the minimal sequences necessary for regulating transcription of the human CYP3A4 gene, more specifically, an element responsive to xenobiotics (the "Xenobiotic Response Element Module” or "XREM” ) and the proximal promoter of the CYP3A4 gene.
  • Xenobiotic Response Element Module the "Xenobiotic Response Element Module” or "XREM”
  • the regulatory nucleic acid molecule of the invention typically contains at least one enhancer capable of regulating transcription of a human CYP3A4 gene when contacted with a nuclear receptor.
  • enhancers are those capable of regulating transcription of a human CYP3A4 gene when contacted with a nuclear receptor bound to a ligand, such as a xenobiotic or steroid.
  • CYP3A4 examples are those capable of regulating transcription of a human CYP3A4 gene when contacted with a nuclear receptor consisting of a heterodimer of PXR (pregnane X receptor, otherwise known as SXR (steroid and xenobiotic receptor) ) and RXR (9-cis retinoic acid receptor) , or CAR (constitutive androstane receptor- ⁇ ) and RXR.
  • PXR pregnane X receptor, otherwise known as SXR (steroid and xenobiotic receptor)
  • RXR (9-cis retinoic acid receptor)
  • CAR constitutitutive androstane receptor- ⁇
  • nucleic acid molecules which have substantially the same nucleotide sequence as a regulatory nucleic acid molecule of the invention would also have sufficient genetic information for reproducing the constitutive and xenobiotic induced tissue specific and developmental expression of the CYP3A4 gene that is observed in a human. Accordingly, it will be understood that nucleotides could be modified or deleted in regions of the regulatory nucleic acid _molecule, more specifically, those regions which do not contain an enhancer such as those described above, without significantly limiting the capacity of the molecule to regulate transcription of the human CYP3A4 gene.
  • the invention provides a non-human mammal of any one of the first to fourth aspects of the invention, further comprising:
  • the further reporter and further regulatory nucleic acid molecules are arranged to permit the further regulatory nucleic acid molecule to regulate transcription of the further reporter nucleic acid molecule.
  • the at least one further regulatory nucleic acid molecule has a sequence shown in SEQ ID NO : 4 . In another embodiment, the at least one further regulatory nucleic acid molecule has a sequence shown in SEQ ID NO : 5.
  • Non-human animals comprising a human PXR or CAR receptor are known. The inventors recognise that the genetic background of these animals could be incorporated into the non-human mammal of the present invention, for example, by conventional breeding techniques.
  • the non-human animal of the invention further comprises at least one human transcription factor for regulating transcription of a human CYP3A4 gene.
  • the transcription factor is a nuclear receptor.
  • the nuclear receptor is a heterodimer of the human PXR (pregnane X receptor, otherwise known as SXR (steroid and xenobiotic receptor) ) and human RXR (9-cis retinoic acid receptor) or human CAR (constitutive androstane receptor- ⁇ ) and human RXR.
  • the reporter nucleic acid molecule can be any molecule which is capable of detection when the reporter nucleic acid molecule is transcribed.
  • the reporter nucleic acid molecule could be the CYP3A4 cytochrome, or the mRNA transcript which is translated to produce the cytochrome.
  • Those reporter molecules which are commercially available, including firefly luciferase, ⁇ - galactosidase, alkaline phosphatase, green fluorescent protein or chloramphenicol acetyl transferase can be used.
  • the reporter nucleic acid molecule is capable of producing a reporter molecule selected from the group of reporter molecules consisting of firefly luciferase, ⁇ -galactosidase, alkaline phosphatase, green fluorescent protein or chloramphenicol acetyl transferase.
  • non-human mammal of the invention is a mouse
  • the inventors believe that any other non-human mammal could be used in the invention, especially those for which standard transgenic techniques have been developed including for example, rat and rabbit.
  • typically the non-human mammal is a mouse.
  • the invention provides a tissue of a non-human mammal of the invention.
  • the tissue is an embryo capable of producing a non-human mammal of the invention.
  • the invention provides a method of determining whether a compound is capable of effecting the transcription of a human CYP3A4 gene the method comprising the following steps:
  • the production of the reporter molecule indicates that the binding compound is capable of effecting the transcription of the human CYP3A4 gene.
  • Any compound can be tested in the method however, preferred compounds are xenobiotic or steroid compounds.
  • FIGURES Figure 1 BRIEF DESCRIPTION OF THE FIGURES Figure 1.
  • the upstream regions of the human CYP3A4 gene are depicted as open boxes with the position of the XREM at approximately -7.5kb of the CYP3A4 gene indicated by cross-hatching.
  • the 5' -flanking region extended from 56bp downstream of the transcription initiation site to a HindiII site at -3,213 in the construct designated - 3CYP3A4/lacZ and to a Kpnl site at -12,926 kb in construct -13CYP3A4/lacZ.
  • the coding region of the E.coli lacZ gene together with eukaryotic translational initiation and termination signals, transcription termination and poly adenylation sites are indicated by a solid box.
  • FIG. 1 Xenobiotic induction of hepatic transgene expression.
  • Female mice from line 9/4 harbouring the - 13CYP3A4/IacZ transgene were treated with various reagents. Histochemical staining of liver slices with X-gal revealed an increased zone of blue staining cells containing ⁇ - galactosidase after treatment with rifampicin, phenobarbital and pregnenolone 16 ⁇ -carbonitrile compared with corn oil treated mice.
  • FIG. 3 Comparison of the xenobiotic induction profile of the -13CYP3A4/lacZ transgene with the mouse Cyp3all gene.
  • Transgenic mice from line 9/4 were treated with a range of xenobiotic reagents and naturally occurring steroids.
  • A. Transgene expression was assessed by determining ⁇ -galactosidase activity in total liver lysates using the ONPG assay. The units of ⁇ -galactosidase activity are given as A 420 /mg liver/minute .
  • Dexamethasone and pregnenolone 16 ⁇ -carbonitrile were the most potent xenobiotic activators of the -13CYP3A4/lacZ transgene, while rifampicin treatment resulted in relatively low levels.
  • the steroids pregnenolone and 17 ⁇ -progesterone were very weak inducers .
  • B. Hepatic expression of the endogenous mouse Cyp3all gene was examined in the same mice by Northern analysis . A similar pattern of induction to the CYP3A4/2acZ transgene was observed with both xenobiotic and endogenous regulators. The data are presented as the mean +/- the standard deviation for 3 animals. Figure 4.
  • Figure 7 The "Xenobiotic-Responsive Enhancer Module” (XREM) of the human CYP3A4 gene. This region encompasses -7836 to -7207 base pairs relative to the transcription initiation site of the CYP3A4 gene.
  • Figure 8. (SEQ ID NO: 4) The 5 '-flanking region of the • human CYP3A7 gene (Genbank Accession No. AF329900) . The extent of the sequences is -11,133 to +52 base relative to the transcription initiation site of the CYP3A7 gene.
  • Figure 9 (SEQ ID NO: 5) Sequence of the 5 '-flanking region of the human MDRl gene (p-glycoprotein gene) encompassing - 10,000 to +200 base pairs relative to the transcription initiation site of the MDRl gene. Sequence derived from within Genbank sequence Accession Number AC002457.
  • Transgene constructs Two transgene constructs were synthesized with the upstream 5' flank of the human cytochrome P450 CYP3A4 gene linked to the E. coli lacZ reporter gene ( Figure 1) .
  • the first construct designated -3CYP3A4/2acZ, contained the region of the CYP3A4 gene from the HindiII site at -3213bp relative to the transcription start site to nucleotide +56bp downstream of the transcription start site.
  • the other construct, designated -13CYP3A4/lacZ included the region of the CYP3A4 gene from the Kpnl site at -12,926bp upstream to +56bp downstream of the transcription start site. It includes the DNA sequences of the XREM region located between -7836 and -
  • the DNA sequence of the CYP3A4 gene between -10468bp and +906bp has been determined and deposited with the GenBank/EMBL/DDJB database under accession number AF185589. Additional sequence information covering the region - 10,469bp to -12,926bp was obtained from publically accessible Genbank files.
  • the E.coli lacZ reporter gene comprises the coding region for the bacterial enzyme ⁇ - galactosidase flanked by DNA sequences for eukaryotic translational start and stop signals, SV40 transcriptional termination and polyadenylation signals and an intron.
  • the CYP3A4/lacZ transgene constructs were released from vector sequences and purified on agarose gels prior to microinj ection
  • mice carrying the CYP3A4/IacZ transgenes were created by microinjection of the DNA constructs into the pro-nuclei of zygotes harvested from FVB/N strain mice. Microinjection and manipulation of embryos were carried by standard techniques. Stable transgenic mouse lines were established by breeding from transgenic founders identified by Southern analysis. Administration of xenobiotics to mice. 8-10 week old male and female mice hemizygous for the -3CYP3A4/2acZ and -
  • 13CYP3A4/IacZ transgenes were used to test the ability of a range of xenobiotics and hormones to activate expression of transgene-derived ⁇ -galactosidase.
  • Mice were administered the following reagents and vehicles by single daily intraperitoneal injection for 4 days: rifampicin/corn oil; dexamethasone phosphate/H 2 0; pregnenolone 16 ⁇ - carbonitrile/2% Tween 20 in H 2 0; phenobarbital/H 2 0; clotrimazole/2% Tween 20; phenytoin/2% Tween 20; 17 -0H progesterone/2% Tween 20; pregnenolone/2% Tween 20.
  • ⁇ - galactosidase activity was visualised in slices and frozen sections of liver and other tissues by staining with X-gal (5-bromo-4-chloro-3-indolyl- ⁇ -D-galactopyranoside) .
  • Tissues were fixed in 0.25% glutaraldehyde, 0. IM phosphate buffer pH7.3, 5mM EGTA, 2 mM MgCl 2 : washed in 0.
  • IM phosphate buffer pH7.3 0.01%sodium deoxycholate, 0.025% NP40, 2mM MgCl 2 and stained by incubation at 37°C in wash solution supplemented with 1mg/ml X-gal, 5mM potassium ferricyanide, and 5mM potassium ferrocyanide .
  • the level of ⁇ -galactosidase activity was determined in whole liver homogenates [lOOmg fresh tissue/ml 0.25M Tris-HCl (pH 7.3)] using the O-nitrophenyl- ⁇ -D-galactopyranoside (ONPG) assay according to standard techniques .
  • ⁇ -galactosidase assay reagent 0.1M sodium phosphate buffer (pH7.3)/lmM MgCl 2 /50 mmol ⁇ -mercaptoethanol/0.88mg/ml ONPG
  • IM Na 2 C0 3 The units of ⁇ -galactosidase activity are given as A 2o /mg liver/minute.
  • transgenic lines were generated with the construct containing the -3.2kb region of the human CYP3A4 gene linked to 2acZ.
  • Transgene-derived ⁇ -galactosidase activity was not detected in kidney, large and small intestine, spleen, lung and liver tissue from mice for all 4 - 3CYP3A4/2acZ transgenic lines treated with vehicle or xenobiotics (Table 1) .
  • transgene expression was readily detected in 3 of the 4 lines carrying the - 13CYP3A4/lacZ construct.
  • Line 9/4 had a very low constitutive level in the liver, with ⁇ -galactosidase detected only in isolated hepatocytes adjacent to major blood vessels.
  • the relative degree of induction for a range of xenobiotics was analysed by determining the transgenic ⁇ -galactosidase activity in liver lysates of mice from line 9/4 ( Figure 3A) .
  • Dexamethasone and pregnenolone 16 ⁇ -carbonitrile were the most potent inducers, while rifampicin activated the transgene to relatively modest levels.
  • Phenobarbital , clotrimazole and phenytoin were intermediate inducers.
  • the induction profile of the transgene in line 9/4 was similar to that observed for the endogenous Cyp3all gene in the same mice (Fig 3B) , likely reflecting the activation profile of the mouse rather than the human PXR.
  • Cyp3all mRNA is only just detectable in males of the FVB/N strain of mice, it may be attributed to the relatively greater degree of induction of the mouse Cyp3all gene in males compared to females ( Figure 3B) .
  • the other line which showed significant transgene expression - 15/10 had a higher constitutive level in both the liver and small intestine in untreated mice. Expression was not detected in other organs, confirming the tissue specificity observed in line 9/4.
  • the same set of reagents were capable of increasing hepatic and intestinal transgene expression to the same levels as in mice from line 9/4. However, the overall degree of induction was not as great as observed in line 9/4 due to the higher basal level in line 15/10.
  • the induction profile was similar with dexamethasone being the most potent activator and rifampicin the least (data not shown) .

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Abstract

La présente invention concerne la génération d'animaux transgéniques non humains comprenant une construction de rapporteur en vue de produire une quantité détectable d'une molécule rapporteur fonctionnellement liée à une molécule d'acide nucléique de régulation transcriptionnelle provenant du gène humain CYP3A4 situé entre le début du site de transcription du gène et un endroit situé 13 000 nucléotides en amont du site. Cette invention concerne également l'utilisation de ces animaux pour déterminer l'effet d'un composé, notamment mais pas uniquement, d'un xénobiotique ou d'un stéroïde, sur la régulation de l'expression du gène CYP3A4 chez l'homme.
PCT/AU2001/001407 2000-11-01 2001-11-01 Animaux transgeniques utilises pour analyser la regulation du gene p450 du cytochrome cyp3a4 Ceased WO2002036784A1 (fr)

Priority Applications (8)

Application Number Priority Date Filing Date Title
CA2426959A CA2426959C (fr) 2000-11-01 2001-11-01 Animaux transgeniques utilises pour analyser la regulation du gene p450 du cytochrome cyp3a4
AU2002213660A AU2002213660A1 (en) 2000-11-01 2001-11-01 Transgenic animals for analysing CYP3A4 cytochrome P450 gene regulation
US10/415,607 US7531712B2 (en) 2000-11-01 2001-11-01 P450 gene regulation
JP2002539530A JP2004512054A (ja) 2000-11-01 2001-11-01 Ｃｙｐ３ａ４シトクロムｐ４５０遺伝子調節を分析するためのトランスジェニック動物
EP01981959A EP1337650A4 (fr) 2000-11-01 2001-11-01 Animaux transgeniques utilises pour analyser la regulation du gene p450 du cytochrome cyp3a4
US12/430,672 US20090307785A1 (en) 2000-11-01 2009-04-27 Transgenic animals for analyzing CYP3A4 cytochrome P450 gene regulation
US12/463,246 US8088968B2 (en) 2000-11-01 2009-05-08 Transgenic animals for analyzing CYP3A4 cytochrome P450 gene regulation
US12/855,383 US8629318B2 (en) 2000-11-01 2010-08-12 Transgenic animals for analyzing CYP3A4 cytochrome P450 gene regulation

Applications Claiming Priority (4)

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AUPR1161 2000-11-01
AUPR1161A AUPR116100A0 (en) 2000-11-01 2000-11-01 P450 gene regulation
AUPR4901A AUPR490101A0 (en) 2001-05-10 2001-05-10 P450 gene regulation ii
AUPR4901 2001-05-10

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US10415607 A-371-Of-International 2001-11-01
US12/430,672 Division US20090307785A1 (en) 2000-11-01 2009-04-27 Transgenic animals for analyzing CYP3A4 cytochrome P450 gene regulation
US12/463,246 Continuation US8088968B2 (en) 2000-11-01 2009-05-08 Transgenic animals for analyzing CYP3A4 cytochrome P450 gene regulation

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002088305A3 (fr) * 2001-04-12 2003-12-11 Xenogen Corp Isolation et identification d'elements regulateurs de transcription murins et humains associes a l'expression de cytochromes
GB2391231B (en) * 2002-07-12 2005-12-14 Imp Cancer Res Tech Modulation of cytochrome P450 reductase activity
WO2007046665A1 (fr) * 2005-10-21 2007-04-26 Samsung Electronics Co., Ltd. Codeur externe et procede de codage externe
US7531712B2 (en) 2000-11-01 2009-05-12 The University Of Syndey P450 gene regulation
US7638614B2 (en) 1998-05-21 2009-12-29 The University Of Sydney Xenobiotic related induction of gene expression
US7700822B2 (en) 2002-07-12 2010-04-20 Cancer Research Technology Limited Modulation of cytochrome P450 reductase activity
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US7638614B2 (en) 1998-05-21 2009-12-29 The University Of Sydney Xenobiotic related induction of gene expression
US7531712B2 (en) 2000-11-01 2009-05-12 The University Of Syndey P450 gene regulation
US8088968B2 (en) 2000-11-01 2012-01-03 The University Of Sydney Transgenic animals for analyzing CYP3A4 cytochrome P450 gene regulation
US8629318B2 (en) 2000-11-01 2014-01-14 University Of Sydney Transgenic animals for analyzing CYP3A4 cytochrome P450 gene regulation
WO2002088305A3 (fr) * 2001-04-12 2003-12-11 Xenogen Corp Isolation et identification d'elements regulateurs de transcription murins et humains associes a l'expression de cytochromes
EP1390744A4 (fr) * 2001-04-12 2006-07-26 Xenogen Corp Isolation et identification d'elements regulateurs de transcription murins et humains associes a l'expression de cytochromes
US7449567B2 (en) 2001-04-12 2008-11-11 Xenogen Corporation Isolation and identification of mouse and human transcription control elements associated with cytochrome expression
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US7700822B2 (en) 2002-07-12 2010-04-20 Cancer Research Technology Limited Modulation of cytochrome P450 reductase activity
US8212105B2 (en) 2004-12-13 2012-07-03 Iti Life Sciences Transgenic mice for assessing drug metabolism and toxicity
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JP5543771B2 (ja) 2014-07-09
AU2007249083B2 (en) 2010-12-16
AU2007249083A1 (en) 2008-01-10
EP1337650A1 (fr) 2003-08-27
AU2007249083B8 (en) 2011-02-03
JP2004512054A (ja) 2004-04-22
CA2426959A1 (fr) 2002-05-10
EP1337650A4 (fr) 2005-12-21
CA2426959C (fr) 2011-10-04
JP2010148508A (ja) 2010-07-08

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