[go: up one dir, main page]

WO2002033054A1 - ANIMAUX KNOCKOUT POLYβ2 - Google Patents

ANIMAUX KNOCKOUT POLYβ2 Download PDF

Info

Publication number
WO2002033054A1
WO2002033054A1 PCT/JP2001/009204 JP0109204W WO0233054A1 WO 2002033054 A1 WO2002033054 A1 WO 2002033054A1 JP 0109204 W JP0109204 W JP 0109204W WO 0233054 A1 WO0233054 A1 WO 0233054A1
Authority
WO
WIPO (PCT)
Prior art keywords
poly
gene
mice
cells
mouse
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/JP2001/009204
Other languages
English (en)
Japanese (ja)
Inventor
Kyoji Ikeda
Noboru Motoyama
Hiroyuki Takai
Yosuke Kobayashi
Miho Watanabe
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.)
Chugai Pharmaceutical Co Ltd
Original Assignee
Chugai Pharmaceutical Co 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 Chugai Pharmaceutical Co Ltd filed Critical Chugai Pharmaceutical Co Ltd
Priority to AU2001295982A priority Critical patent/AU2001295982A1/en
Priority to JP2002536424A priority patent/JPWO2002033054A1/ja
Publication of WO2002033054A1 publication Critical patent/WO2002033054A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • 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
    • A01K67/0276—Knock-out vertebrates
    • 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
    • A01K2217/07—Animals genetically altered by homologous recombination
    • A01K2217/075—Animals genetically altered by homologous recombination inducing loss of function, i.e. knock out
    • 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

Definitions

  • the present invention relates to cells and rodents in which the poly-2 gene has been knocked out, and to uses thereof.
  • the DNA polymerase-2 (poly-2) gene contains a BRCT motif at the N-terminus and has a region at the C-terminus that shows about 30% homology with DNA polymerase (Nucleic Acids Research , 28, 3684-3693, (2000) , Journal of Molecular Biology, 301, 851-867, (2000), 'GenBank AF176098, DDBJ Keio 1019, EMBL AF176097) o, however, not been elucidated with respect to its function However, an experimental system useful for elucidating its function is required.
  • the poly-2 gene pair is inactivated and there is a genetically modified animal that has no poly52 activity, a cell line established from the genetically modified animal (genetically modified mouse-derived cell line), or an ES cell line, the individual This makes it possible to elucidate the physiological functions at the level of cells and at the cell level, and it can be used for research on therapeutics for diseases related to the gene. Disclosure of the invention
  • the present invention provides a genetically modified animal in which poly-2 gene expression is artificially suppressed, a cell line established from the genetically modified animal, a gene-modified ES cell line, and uses thereof.
  • the purpose is to:
  • the present inventors have conducted intensive studies in order to solve the above problems, and as a result, established ES cells in which one of the loci was inactivated by using gene targeting, and used the cells to develop ES cells.
  • Mera mice were produced, and by crossing using the chimeric mice, a genetically modified animal in which one or both of the poly; 52 gene pair was inactivated was successfully produced.
  • Genetically modified animals in which both gene pairs of the Poly ⁇ 2 gene have been inactivated include hydrocephalus, poor growth, visceral inversion, poor sperm maturation, cilia structure in the pilus of ventricular ependymal cells and tracheal epithelial cells It was characterized by abnormalities (dynein inner arm defect). Therefore, ES cells and animals in which the poly / 52 gene has been inactivated are effective in elucidating these diseases and developing therapeutic agents.
  • the present invention relates to a genetically modified animal and an ES cell line in which the expression of the poly ⁇ 2 gene is artificially suppressed, a cell line established from the genetically modified animal, and the use thereof.
  • the genetically modified ES cells and genetically modified animals of the present invention are characterized in that poly-2 gene expression is artificially suppressed.
  • the mouse poly /? 2 gene consisted of eight exons and was encoded on chromosome 19 ( Figure 1). In adult mice, poly 2 mRNA was expressed in almost all tissues, but the highest expression was observed in testis. poly 22 was localized in the nucleus as a protein with a molecular weight of about 60 kDa.
  • Figure 2 shows a comparison of the mouse and human poly-2 gene sequences.
  • the target animal for the modification of the poly52 gene is preferably a rodent such as a mouse, rat or hamster, and among them, a mouse is particularly preferable.
  • ES cells targeted for poly-2 gene modification are also preferably derived from rodents, and are particularly preferably derived from mice.
  • the expression of the poly-2 gene is suppressed means that the expression of the poly-2 gene is completely suppressed, or that the expression of only one gene of the gene pair is suppressed. It is intended to include the case where it is imperfectly suppressed, such as the case where In the present invention, it is preferable that the expression of the poly-2 gene is specifically suppressed.
  • a method of deleting the poly ⁇ 2 gene or a part of the expression control region of the poly ⁇ 2 gene may be used.
  • Examples of the method include a method for deleting the gene, and a method for inactivating the poly-2 gene by inserting a foreign gene into one or both of the gene pairs of the poly-2 gene is preferable. That is, in a preferred embodiment of the present invention, the genetically modified ES cell and the genetically modified animal are characterized in that a foreign gene has been inserted into one or both of the poly / -2 gene pair.
  • the genetically modified mouse of the present invention can be prepared as follows. First, DNA containing the exon portion of the poly-2 gene is isolated from a mouse, and an appropriate marker gene is inserted into this DNA fragment to construct a targeting vector. This evening targeting vector is introduced into a mouse ES cell line by an electroporation method or the like, and a cell line in which homologous recombination has occurred is selected.
  • a marker gene to be inserted an antibiotic resistance gene such as a neomycin resistance gene is preferable.
  • a cell line in which homologous recombination has occurred can be selected only by culturing in a medium containing the antibiotic.
  • the thymidine kinase gene can be linked in the evening. You. As a result, cell lines that have undergone heterologous recombination can be excluded. In addition, a diphtheria toxin A fragment (DT-A) gene or the like can be linked to the getter vector in the evening. As a result, cell lines that have undergone heterologous recombination can be excluded. In addition, a homologous recombinant can be assayed by PCR and Southern blot to efficiently obtain a cell line in which one of the gene pairs of the poly52 gene has been inactivated.
  • DT-A diphtheria toxin A fragment
  • a cell line in which homologous recombination has occurred it is preferable to create a chimera using a plurality of clones, since there is a risk of unknown gene disruption due to gene insertion in addition to the homologous recombination site.
  • the obtained ES cell line is injected into mouse blastocysts to obtain chimeric mice.
  • a mouse in which one of the gene pairs of the poly-2 gene has been inactivated can be obtained.
  • a mouse in which both gene pairs of the poly52 gene have been inactivated can be obtained.
  • Gene modification can also be performed in animals other than mice, in which ES cells have been established, by the same method.
  • An ES cell line in which both the poly-2 gene pair has been inactivated can also be obtained by the following method. That is, by culturing an ES cell line in which one of the gene pairs has been inactivated in a medium containing a high concentration of antibiotics, a cell line in which the other of the gene pair has also been inactivated, that is, the poly-2 gene, An ES cell line in which both gene pairs have been inactivated can be obtained. Alternatively, it can also be prepared by selecting an ES cell line in which one of the gene pairs has been inactivated, introducing the gettering vector again into this cell line, and selecting a cell line in which homologous recombination has occurred. it can. It is preferable to use a marker gene to be inserted into the evening-getting vector, which is different from the marker gene described above.
  • a known method can be used.
  • the method of primary culture of fetal cells can be used (Shinsei Kagaku Jikken Koza, Vol. 18, p. 125-129, Tokyo Chemical Dojin, and Ma Mouse embryo operation manual, pp. 262-264, Modern Publishing).
  • mice in which both gene pairs were inactivated were considered to have hydrocephalus. In addition, some mice exhibited visceral inversion. Furthermore, in female mice, an increase in immature sperm cells and a decrease in mature sperm cells were observed. In addition, dynein inner arm deficiency was observed in fimbriae of ependymal cells and tracheal epithelial cells. No abnormalities were found in the differentiation of T and B lymphocytes in the thymus and spleen.
  • mice in which both gene pairs of the poly-2 gene were inactivated showed hydrocephalus, poor growth, visceral inversion, poor sperm maturation, fimbriae of ependymal cells and tracheal epithelial cells.
  • mice in which both gene pairs of the poly-2 gene were inactivated showed hydrocephalus, poor growth, visceral inversion, poor sperm maturation, fimbriae of ependymal cells and tracheal epithelial cells.
  • the poly-2 gene is thought to be involved in hydrocephalus, poor growth, visceral inversion, poor sperm maturation, dynein inner arm deficiency in fimbria of ventricular ependymal cells and tracheal epithelial cells.
  • poly-2 has immotile cilia syndrome, such as total visceral inversion with bronchiectasis and chronic sinusitis, impaired ciliary movement, impaired ciliary mucus transport in respiratory airway epithelium, etc. Involvement in the characteristic Kartagener's syndrome is suggested.
  • the genetically modified animal of the present invention can be used not only for analyzing the detailed function of the polyZ gene, but also for hydrocephalus, poor growth, visceral position, It can be used to elucidate diseases such as poor sperm maturation and i-painted otile cilia syndrome. Furthermore, it can be used for screening of therapeutic agents for those diseases.
  • poly-2 gene and protein can be used as therapeutics for the above diseases, etc.
  • poly ⁇ 2 gene is expected to be used for the treatment of the above diseases by gene therapy. You. Brief description of the drawings ''
  • FIG. 1 shows the exon 'intron structure of the mouse poly / 52 gene.
  • A shows the overall image of the exon-intron structure
  • B shows the nucleotide sequence of the boundary region between the exon and the intron.
  • FIG. 2 shows a comparison of the nucleotide sequences of the mouse and human poly / 52 genes.
  • FIG. 4 is a photograph showing a section of the brain of a poly52 + / + mouse and a poly / 52 ⁇ / ⁇ mouse.
  • Embryo 14.5 is a photograph showing a cross-section of the brain of poly /? 2 + / + mice (left) and poly 52-/-mice (right).
  • Figure 5 is a photograph showing a serial sagittal section of the brain.
  • FIG. 4g The ventricular ependymal cells of the poly? 2 + / + mouse make up several layers of cells surrounding the ventricle (Fig. 4g, h), but in the brain of the poly52-/-mouse (right). Hyperplasia of ependymal cells in the third ventricle (3V) and middle cerebral aqueduct (Aq) is observed, and the layer structure is broken (Fig. 4c-f). As can be seen from the enlarged photograph inserted in c, at the hyperplastic site of the ependymal cells, there are mononuclear cells with a spherical, well-defined nucleus and mononuclear cells with eosinphilic cytoplasm (Fig. 4c).
  • FIG. 6 is a photograph showing the result of examining the localization of a transcript (mRNA) of the poly32 gene in the brain and testis by in situ hybridization.
  • mRNA transcript
  • a, b Photographs of a sperm obtained from the epididymis tail observed with a phase contrast microscope. Compared to the shape of spermatozoa of 9-week-old poly? 2 + / + mice (a), the spermatozoa of poly /? 2-/-mice (b) have short tails, those without tails, Malformed spermatozoa such as those with a circular head were seen.
  • c-h are photographs showing the results of pathological analysis of testis.
  • the poly? 2-/-mouse testis (d) has the same tubule structure as the poly / 52 + ⁇ mouse testis (c).
  • 3 is a photograph showing the results of pathological analysis of the epididymis body part. .
  • Poly 32 + / + mice (i) show mature sperm cells, whereas poly /? 2-/-mice (j) have few mature sperm cells, immature sperm cells and eosinophils Many substances were found.
  • FIG. 8 shows the survival curve of poly 2 ⁇ / ⁇ mice.
  • FIG. 4 presents photographs showing the results of Northern blot analysis of the expression of poly ⁇ Z gene transcript (mRNA) in testis of 'c. poly? 2-/-mice. EF-1 was used as a control. .
  • FIG. 10 is an anatomical photograph showing the internal organ arrangement of poly-2 ⁇ / ⁇ and poly-2 + / + mice. Ribs and liver have been resected for better visibility of internal organs. The internal organs of poly ⁇ 2 ⁇ / ⁇ mice are mirror images of the internal organs of poly 2 + / + mice. c In the figure, Ap is Apex (apical), Fs is Forestomach (foregoma), and Sp is Spleen. (Spleen).
  • FIG. 11 is a photograph of the pilus structure of a poly-2 /-mouse observed with an electron microscope.
  • the upper part shows the cross section of the fimbria of the ependymal cells of the ventricle, and the lower part shows the cross section of the pili of the tracheal epithelial cells.
  • 2- /-mice dynein inner arm deficiency is found in the pili of ependymal cells (upper right) and the pili of tracheal epithelial cells (lower right).
  • Figure 12 is a schematic diagram of two dynein arms (outer arm and inner arm) of a double microtubule.
  • the mouse poly52 genomic gene was cloned.
  • plaque hybridization was performed using mouse poly-2c DNA as a probe, and a genomic DNA fragment of approximately 20kb containing poly / -2 gene exon 1 was isolated. The site was mapped.
  • exon 1 to exon 6 of the poly-2 gene with the neomycin resistance gene expression cassette by homologous recombination include the gene fragment containing the 5 'side sequence of exon 1 and exons 7 and 8 at both ends of the neomycin resistance gene expression cassette
  • the homologous region was arranged by ligating the gene fragments, and the tk expression cassette was further ligated to the 3 'downstream to create a homologous recombination vector (FIG. 9a, upper panel).
  • the poly-2 locus has a restriction enzyme Xbal recognition site (X) upstream of exon 1 and a restriction enzyme Spel recognition site (Sp) between exons 3 and 4 (middle of Fig. 9a).
  • the homologous recombination vector was introduced into mouse ES cell E14 (Hopper, M. et al .: Nature, 326, 292-295, 1987) by electroporation, followed by G418 and ganciclovir. Selection culture was performed. The obtained G418Z ganciclovir-resistant colonies were tested for homologous recombinants by PCR and Southern blotting, and four cell lines in which the mutant poly 2 gene was inserted into one of the poly / -2 gene pairs were obtained.
  • the resulting clones were confirmed by Southern analysis for insertion of the mutant gene (FIG. 9b).
  • the mutant gene was not inserted, only the llkb band was detected, whereas the cell lines 96, 97, and 110 obtained by introducing the vector for homologous recombination and selecting the drug were used.
  • a llkb band derived from the normal poly-2 locus and a 9.2-kb band derived from the mutant poly / -2 locus were detected.
  • ES cells in which one of the poly ⁇ 2 genes was inactivated were injected into blastocysts derived from C57BL / 6 mice to obtain chimeric mice.
  • a chimeric mouse prepared using ES cells in which one of the poly-2 gene pairs has been inactivated is crossed with C57BL / 6 mice, and one of the poly-2 genes pairs consists of inactivated cells
  • a mouse (hereinafter, referred to as "poly? 2 +/- mouse") was obtained.
  • poly? 2-/-mice mice in which both poly ⁇ 2 gene pairs were inactivated
  • mice with marked lateral ventricular dilation show apoptosis in ependymal cells around the ventricle. Such apoptosis is not observed around the ventricle before ventricular dilation.
  • Ventricular dilatation is seen in the lateral and third ventricles, but not in the fourth and middle ventricles.
  • hyperplasia of ependymal cells in the third ventricle and the middle cerebral aqueduct is observed from day 14.5 onward (Fig. 4c-e).
  • the ventricular ependymal cells comprise several layers of cells that surround the ventricle (Fig. 5g, h).
  • hyperplasia of the ependymal cells of the third ventricle and the middle cerebral aqueduct was observed, and the layer structure was disrupted (Fig. 5 af).
  • mononuclear cells with a spherical and well-defined nucleus and mononuclear cells with eosinphilic cytoplasm are observed (Fig. 5c).
  • Electron microscopy of ependymal cells showing hydrocephalus reveals multilayering of ependymal cells and disruption of the microvilli margin (Fig. 4f).
  • mice in which both poly-2 genes are inactivated is due to cerebrospinal fluid production and / or reabsorption due to occlusion of the cerebrospinal fluid circulation and disruption of ependymal microvillous margins. It is considered abnormal.
  • Poly ⁇ 2-/-mice with hydrocephalus show growth retardation compared to wild-type mice in which the poly ⁇ 2 gene is not inactivated (hereinafter referred to as "poly52 + / + mice”). 3 a). On days 14 and 21 after birth, poly? 2-/-mice weighed poly 2 + / + About 60-80% of mice.
  • poly ⁇ 2-/-mice begin to die at 3 weeks of age and have a 50% survival rate by 5 weeks of age (Figure 8).
  • the dead poly 2 ⁇ / ⁇ mice are mostly individuals with hydrocephalus, and the poly 52 ⁇ / ⁇ mice surviving beyond 5 weeks of age are apparently identical to normal mice.
  • poly-2 mRNA The localization of the transcript (mRNA) of the poly52 gene was examined by in situ and hybridization.
  • poly-2 mRNA is localized in cells around the ventricle of the fetal brain of wild-type mice and in adult ependymal ependymal cells and choroid plexus.In poly52-/-mice, poly-2 mRNA is expressed in those cells. No localization is observed ( Figures 6a-c).
  • the localization of the transcript (mRNA) of the poly /? 2 gene was examined by in situ hybridization. Poly-2mMA is localized in the testes of wild-type mice. In poly / 52-/-mice, no localization of poly-2 mRNA in testis was observed (Fig. 6d). .
  • RNA poly-2 transcripts
  • Fig. 9c The expression of poly-2 transcripts (mRNA) in testis of poly-2 /-mice was analyzed by Northern blot analysis (Fig. 9c). In poly? 2 + / + mice and (poly? 2 +/- mice, the expression of poly /? 2 gene mRNA is observed, but in poly /? 2-/-mice, the expression of poly? 2 gene is observed.
  • the lower panel in Fig. 9c shows the expression of EF-1 cells, which demonstrates the presence of the RNA sample in each lane.
  • Dyskinesia due to ciliary structural abnormalities similar to those found in cilia of tracheal epithelial cells and ventricular ependymal cells also occur in the cilia of nasal mucociliary cells that make up the nasal mucosa It is presumed that rhinitis has developed.
  • ES cells and mice in which poly-2 gene expression is artificially suppressed wherein the mice have hydrocephalus, poor growth, visceral inversion, poor sperm maturation, ventricular ependymal cells and It was found to be characterized by abnormal ciliary structure (dynein inner arm defect) in the pili of tracheal epithelial cells. Therefore, if the genetically modified ES cells, genetically modified animals, or cell lines established from the animals of the present invention are used, the function of the poly /? 2 gene can be analyzed, as well as hydrocephalus, poor growth, visceral inversion, and sperm.
  • poly-2 gene and protein can be used as therapeutic agents for the above-mentioned diseases and the like.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Environmental Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Animal Behavior & Ethology (AREA)
  • Animal Husbandry (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Molecular Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention concerne des animaux génétiquement modifiés chez lesquels une ou les deux paires génétiques de gènes polyβ2 sont inactivées. Lesdits animaux sont créés par établissement de cellules souches embryonnaires dans lesquels un des locus de gène est inactivé au moyen de la méthode de ciblage génétique, création de souris chimériques au moyen desdites cellules, et conjugaison des souris chimériques ainsi obtenues. Lesdits souris knockout sont caractérisées en ce qu'elles souffrent d'hypocéphalie, d'hypoplasie, d'inversion viscérale, de défaut de maturité spermatique et de défaut structurel ciliaire dans des cellules pileuses et épendymaires et des cellules épithéliailes trachéales (bras intérieur de la dynéine manquant).
PCT/JP2001/009204 2000-10-20 2001-10-19 ANIMAUX KNOCKOUT POLYβ2 Ceased WO2002033054A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU2001295982A AU2001295982A1 (en) 2000-10-20 2001-10-19 Polybeta2 knockout animals
JP2002536424A JPWO2002033054A1 (ja) 2000-10-20 2001-10-19 ｐｏｌｙβ２遺伝子欠損動物

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
JP2000-321714 2000-10-20
JP2000321714 2000-10-20
JP2000-372555 2000-12-07
JP2000372555 2000-12-07
JP2001120294 2001-04-18
JP2001-120294 2001-04-18

Publications (1)

Publication Number Publication Date
WO2002033054A1 true WO2002033054A1 (fr) 2002-04-25

Family

ID=27345002

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2001/009204 Ceased WO2002033054A1 (fr) 2000-10-20 2001-10-19 ANIMAUX KNOCKOUT POLYβ2

Country Status (3)

Country Link
JP (1) JPWO2002033054A1 (fr)
AU (1) AU2001295982A1 (fr)
WO (1) WO2002033054A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005017155A1 (fr) 2003-06-18 2005-02-24 Chugai Seiyaku Kabushiki Kaisha Transporteur du fucose
EP2208783A1 (fr) 2004-12-22 2010-07-21 Chugai Seiyaku Kabushiki Kaisha Procédé de préparation d'un anticorps à l'aide d'une cellule dont la fonction de transporteur du fucose est inhibée
US7829757B2 (en) 2001-10-24 2010-11-09 Chugai Seiyaku Kabushiki Kaisha SGRF gene-modified mouse

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001025442A1 (fr) * 1999-10-01 2001-04-12 Consejo Superior De Investigaciones Cientificas Polymerase lambda d'adn et ses utilisations

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001025442A1 (fr) * 1999-10-01 2001-04-12 Consejo Superior De Investigaciones Cientificas Polymerase lambda d'adn et ses utilisations

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
AOUFOUCHI S. ET AL.: "Two novel human and mouse DNA polymerases of the polX family", NUCLEIC ACIDS RES., vol. 28, no. 18, 15 September 2000 (2000-09-15), pages 3684 - 3693, XP002157837 *
LINDEMAN G.J. ET AL.: "A specific, nonproliferative role for E2F-5 in choroid plexus function revealed by gene targeting", GENES DEV., vol. 12, no. 8, 1998, pages 1092 - 1098, XP002907709 *
NAGASAWA K. ET AL.: "Identification and characterization of human DNA polymerase beta2, a DNA polymerase beta-related enzyme", J. BIOL. CHEM., vol. 275, no. 40, 6 October 2000 (2000-10-06), pages 31233 - 31238, XP001011150 *
TAKAI H. ET AL.: "Aberrant cell cycle checkpoint function and early embryonic death in Chk1(-/-) mice", GENES DEV., vol. 14, no. 12, 15 June 2000 (2000-06-15), pages 1439 - 1447, XP002907710 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7829757B2 (en) 2001-10-24 2010-11-09 Chugai Seiyaku Kabushiki Kaisha SGRF gene-modified mouse
WO2005017155A1 (fr) 2003-06-18 2005-02-24 Chugai Seiyaku Kabushiki Kaisha Transporteur du fucose
EP2228445A1 (fr) 2003-06-18 2010-09-15 Chugai Seiyaku Kabushiki Kaisha Transporteur du fucose
EP2208783A1 (fr) 2004-12-22 2010-07-21 Chugai Seiyaku Kabushiki Kaisha Procédé de préparation d'un anticorps à l'aide d'une cellule dont la fonction de transporteur du fucose est inhibée
EP2216404A1 (fr) 2004-12-22 2010-08-11 Chugai Seiyaku Kabushiki Kaisha Procédé de préparation d'anticorps basé sur l'utilisation d'une cellule dont la fonction de transporteur du fucose est inhibée

Also Published As

Publication number Publication date
JPWO2002033054A1 (ja) 2004-02-26
AU2001295982A1 (en) 2002-04-29

Similar Documents

Publication Publication Date Title
Brody et al. Ciliogenesis and left–right axis defects in forkhead factor HFH-4–null mice
Chen et al. Mutation of the mouse hepatocyte nuclear factor/forkhead homologue 4 gene results in an absence of cilia and random left-right asymmetry.
CN107164406B (zh) 胰岛β细胞条件性敲除Tmem30a基因小鼠模型的构建方法及应用
Cho et al. The Na+-Ca2+ exchanger is essential for embryonic heart development in mice
CN108029638B (zh) 一种视网膜色素变性疾病动物模型的构建方法及应用
Lin et al. Deletion of the aryl hydrocarbon receptor-associated protein 9 leads to cardiac malformation and embryonic lethality
CN109182355B (zh) 视网膜新生血管疾病模型的构建方法和应用
Warren et al. A Sall4 mutant mouse model useful for studying the role of Sall4 in early embryonic development and organogenesis
Lee et al. IFT46 plays an essential role in cilia development
Appelbe et al. Disruption of the mouse Jhy gene causes abnormal ciliary microtubule patterning and juvenile hydrocephalus
US8952214B2 (en) Animal model for chronic obstructive pulmonary disease and cystic fibrosis
WO1998051781A1 (fr) Animal transgenique exprimant une proteine preseniline 1 mutante non native de type sauvage de la maladie d'alzheimer familiale dans un contexte depourvu de preseniline 1 native
WO2002033054A1 (fr) ANIMAUX KNOCKOUT POLYβ2
KR101954726B1 (ko) 펜드리드 증후군 동물 모델 및 이의 제조 방법
US6225525B1 (en) ATP-binding cassette transporter (ABC1) modified transgenic mice
CN115777626B (zh) Ift140基因视杆细胞条件敲除小鼠模型的构建方法及应用
EP1910416B1 (fr) Modele animal non humain pour maladie cardio-vasculaire caracterisee par un gene de fibuline-4 fragmente
CN117230077B (zh) Hakai基因在RP疾病模型构建中的应用及构建方法
JP4303907B2 (ja) ラディキシンノックアウト動物
KR100517831B1 (ko) ＰＳ２ 돌연변이 유전자 및 ＡＰＰｓｗ 유전자를 발현하는 이중 형질전환 치매 마우스
KR100430199B1 (ko) 칼슘 이온 통로 알파１ｄ 유전자 변이 생쥐 및 그의 제조방법
JP4696316B2 (ja) Ｃｄ９／ｃｄ８１二重欠損非ヒト動物
JP2009225697A (ja) Ａｂｃａ１２遺伝子機能欠損マウス
CN117660527A (zh) 一种ABCA7-Floxp小鼠模型的构建方法和应用
CN117551700A (zh) 一种fsgs疾病模型的构建方法及应用

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PH 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: A1

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 GQ GW ML MR NE SN TD TG

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

Ref country code: JP

Ref document number: 2002 536424

Kind code of ref document: A

Format of ref document f/p: F

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase