WO2002097090A1 - Genes with es cell-specific expression - Google Patents
Genes with es cell-specific expression Download PDFInfo
- Publication number
- WO2002097090A1 WO2002097090A1 PCT/JP2002/005350 JP0205350W WO02097090A1 WO 2002097090 A1 WO2002097090 A1 WO 2002097090A1 JP 0205350 W JP0205350 W JP 0205350W WO 02097090 A1 WO02097090 A1 WO 02097090A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cells
- seq
- dna
- protein
- sequence
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6881—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
-
- 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0603—Embryonic cells ; Embryoid bodies
- C12N5/0606—Pluripotent embryonic cells, e.g. embryonic stem cells [ES]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
Definitions
- the present invention relates to an ECAT gene (ES cell associated transcript gene) that is specifically expressed in ES cells (embryonic stem cells) and its use.
- ECAT gene ES cell associated transcript gene
- Embryonic stem (ES) cells are isolated from early mammalian embryos and proliferate semipermanently while maintaining the ability to differentiate into all cells in the body, that is, totipotency. ES cells were first established in mice in 1981, and brought a revolutionary technology of gene function analysis using knockout mice. Furthermore, since the establishment of human ES cells was reported in 1998, application to regenerative medicine has been greatly expected. The aim is to transplant the myocardium and nerve cells differentiated from ES cells into patients with myocardial infarction and neurodegenerative disease to recover their function.
- Cell transplantation therapy as exemplified by bone marrow transplantation for leukemia, has two issues: securing sufficient transplanted cells and suppressing rejection.
- securing sufficient transplanted cells By using semi-permanently dividing ES cells, the problem of securing sufficient cells can be solved at once.
- somatic cell cloning technology rejection can be overcome. If ES cells are established from cloned embryos produced from the patient's somatic cells and used for transplantation, rejection cannot occur because they have the same gene as the patient. Thus, ES cells have the potential to overcome both challenges in cell transplantation therapy.
- ES cells have such great potential, human ES cells are more difficult to establish and maintain than mouse ES cells, and it is necessary to develop reliable establishment and culturing techniques. In addition, embryos must be sacrificed to establish human ES cells. Furthermore, when combined with somatic cell cloning technology, it can be easily linked to the production of cloned humans. In order to solve such ethical problems, there is a demand for the development of a technology for directly producing ES-like cells having totipotency from somatic cells without passing through embryos.
- a gene that plays an important role in the development of these technologies is a gene that is specifically expressed in totipotent cells such as ES cells (ES cell associated transcript gene, hereinafter referred to as EC AT gene).
- the ECAT gene is a marker for determining whether a cell is an ES cell.
- ES cells can be efficiently selected from a mixed culture of many types of cells (Table 9-500004). No. 6,146,888).
- the conversion to ES-like cells can be promoted by inducing the expression of the ECAT gene in somatic cells.
- ECAT The only reported ECAT gene so far is the transcription factor Oct3 (Oct4, also called POU5 ⁇ 1; hereafter Oct_3Z4) gene.
- Oct3 also called POU5 ⁇ 1; hereafter Oct_3Z4 gene.
- hOct_3Z4 gene Takeda et al., Nucleic Acids Res. 20: 4613-4620, 1992.
- hOct_3 / 4 is a transcription specifically expressed in ES cells and EG cells (embryonic germ cells). It is a factor that loses its expression as the cells differentiate, and is used as a marker for ES cells.
- Oct_3Z4 is expressed not only in totipotent cells but also in trophectoderm cells in addition to totipotent cells. (Biol Reprod 63: 1698-1705, 2000), using only the Oct-3 / 4 gene as an index. In order to prevent this risk, it is desirable to identify multiple ECAT genes and use them in combination.
- An object of the present invention is to provide a novel ECAT gene. More specifically, an object of the present invention is to provide a method for screening ES cells using a new ECAT gene and a gene product peptide encoded by the gene, and a probe for ES cell selection.
- hECAT human homologous gene
- an ES cell selection probe comprising any one of the DNAs having the nucleotide sequences set forth in SEQ ID NOs: 1, 3, 4, 5, 6, 7 or 8.
- An ES cell selection probe comprising a DNA encoding a protein that is specifically expressed and a DNA that can hybridize under stringent conditions.
- a probe for selecting an ES cell which comprises at least one of a DNA consisting of the nucleotide sequence of SEQ ID NO: 9, 13, 15, 17, 19, 21, 23, or 41 .
- ES cell selection probe comprising DNA encoding a protein that is specifically expressed.
- the probe for selecting an ES cell according to the above (6) which comprises a DNA encoding a protein expressed in E. coli.
- ES cell selection probe comprising a DNA that can hybridize under mild conditions.
- (9) It consists of a base sequence in which one or several bases are deleted, substituted or added in the base sequence described in SEQ ID NO: 9, 13, 15, 17, 19, 21 or 23, and is used for ES cells.
- the probe for selecting an ES cell according to the above (8) which comprises a DNA encoding a protein that is specifically expressed and a DNA that can hybridize under stringent conditions.
- a probe for selecting ES cells which comprises any one of DNAs comprising the nucleotide sequence of SEQ ID NO: 27, 29, 31, 33, 35, 37 or 43.
- a gene comprising any one of the following DNAs (a) to (c):
- (c) a DNA consisting of a base sequence in which one or several bases have been deleted, substituted or added in the base sequence of (a), and which encodes a protein specifically expressed in ES cells.
- a gene comprising any one of the following DNAs (a) to (c).
- nucleotide sequence comprising a nucleotide sequence in which one or several nucleotides have been deleted, substituted or added in the nucleotide sequence of (a), and encoding a protein specifically expressed in ES cells.
- (c) D comprising a nucleotide sequence in which one or several nucleotides have been deleted, substituted or added in the nucleotide sequence of (a), and encoding a protein specifically expressed in ES cells.
- a protein comprising an amino acid sequence in which one or several amino acids have been deleted, substituted or added in the amino acid sequence of (a), and which is specifically expressed in ES cells (26) DNA consisting of the base sequence described in SEQ ID NO: 9, 13, 15, 17, 19, 21, 23 or 41, or SEQ ID NO: 10, 14, 16, 18, 20, 20, '
- a method for screening ES cells comprising analyzing the expression status of a protein comprising the amino acid sequence described in 22, 24 or 42 in a cell.
- DNA consisting of the nucleotide sequence of SEQ ID NO: 9, 13, 15, 17, 19, 21 or 23, or SEQ ID NO: 10, 14, 16, 18, 20, 22, or 24
- a method for screening ES cells comprising analyzing the state of expression of a protein comprising the amino acid sequence to be expressed in a cell.
- (31) DNA consisting of the base sequence described in SEQ ID NO: 27, 29, 31, 33, 35 or 37, or described in SEQ ID NO: 28, 30, 32, 34, 36 or 38 (30) The method for screening ES cells according to (30) above, wherein the state of expression of the protein comprising the amino acid sequence in the cell is analyzed.
- An ES cell selection probe comprising a continuous partial sequence of 20 bases or more that does not contain a repetitive sequence in the sequence, and comprising a DNA having a sequence specific to a gene specifically expressed in ES cells.
- the present invention relates to a recombinant vector having a DNA encoding a gene specifically expressed in an ES cell or a protein specifically expressed in an ES cell, particularly a betater for forced expression of a differentiation inhibitory gene, and a vector comprising the vector.
- the present invention further provides a recombinant vector comprising a gene specifically expressed in ES cells or a genomic DNA fragment containing DNA encoding a protein specifically expressed in ES cells, and a selection gene such as a drug resistance gene incorporated into a genomic DNA fragment.
- the present invention relates to a vector for selecting ES cells, and a transformed cell transformed with the vector.
- FIG. 1 shows the results of Northern blot analysis of the expression of each ECAT gene in ES cells and 12 kinds of organs of adult mice.
- Figure 2 shows ES cells, mesenchymal stem cells, and ECA in 13 types of human adult organs.
- FIG. 2 is a diagram showing the expression of T gene analyzed by Northern blot.
- the present invention relates to a gene specifically expressed in ES cells (hereinafter, also referred to as ES cell-specific expression gene), that is, an ECAT gene. It is possible to determine whether the cells are ES cells using the presence or absence of the expression of the ECAT gene as an index.
- the present invention provides a probe for selecting ES cells suitable for determining whether such ES cells are correct or not. Specific examples of the probe include a DNA consisting of the nucleotide sequence shown in any one of SEQ ID NOs: 1 to 8, a DNA consisting of the nucleotide sequence shown in SEQ ID NO: 9 (hereinafter, ECAT 1 gene), DNA consisting of the base sequence described in SEQ ID NO: 11 in the sequence listing
- ECAT2 gene (Hereinafter referred to as the ECAT2 gene), consisting of the nucleotide sequence described in SEQ ID NO: 13 in the Sequence Listing.
- ECAT3 gene DNA consisting of the nucleotide sequence shown in SEQ ID NO: 15 in the Sequence Listing (hereinafter ECAT4 gene), DNA consisting of the nucleotide sequence shown in SEQ ID NO: 17 in the Sequence Listing (hereinafter ECAT5 gene) ),
- ECAT6 gene DNA consisting of the base sequence described in SEQ ID NO: 19
- ECAT7 gene DNA consisting of the base sequence described in SEQ ID NO: 21
- ECAT9 gene A polynucleotide comprising a DNA having the described nucleotide sequence (hereinafter referred to as the ECAT8 gene) or a DNA having the nucleotide sequence described in SEQ ID NO: 41 (hereinafter referred to as the ECAT9 gene) is exemplified.
- the ECAT8 gene DNA having the described nucleotide sequence described in SEQ ID NO: 41
- the base sequence described above may have undergone modification such as substitution, deletion or addition.
- a polynucleotide containing a DNA encoding a protein that hybridizes to the ECAT gene under stringent conditions and is specifically expressed in ES cells, or one or several bases in the base sequence of the ECAT gene The present invention also relates to a polynucleotide comprising a DNA comprising a base sequence having deletion, substitution, or addition of DNA and capable of hybridizing under stringent conditions with DNA encoding a protein specifically expressed in ES cells.
- hECAT2 gene a DNA consisting of the nucleotide sequence shown in SEQ ID NO: 27 of the Sequence Listing
- a DNA comprising the sequence hereinafter referred to as hECAT3 gene
- hECAT4 gene a DNA comprising the nucleotide sequence described in SEQ ID NO: 31
- hECAT5 gene DNA consisting of the base sequence described below
- hECAT7 gene DNA consisting of the base sequence shown in SEQ ID NO: 35 in the Sequence Listing
- hECAT8 gene a polynucleotide comprising a DNA comprising the nucleotide sequence described in SEQ ID NO: 43
- hECAT9 gene a polynucleotide comprising a DNA comprising the nucleotide sequence described in SEQ ID NO: 43
- DNA having the nucleotide sequence described in SEQ ID NO: 25 in the Sequence Listing that is, DNA encoding Oct-3Z4, or DNA having the nucleotide sequence described in SEQ ID NO: 39 in the Sequence Listing, ie, hOct-3 /
- a polynucleotide containing DNA encoding 4 can also be used as a probe for ES cell selection.
- the Oct-3 / 4 gene is also expressed in vegetative ectoderm cells, so it preferably contains an ECAT gene other than the Oct-3 / 4 gene or the hOct-3 / 4 gene.
- the novel ES cell selection probe of the present invention such as a polynucleotide, in combination. Further, even in the case of the novel ES cell selection probe containing the ECAT gene, it is preferable to use several types of probes in order to more accurately determine whether or not the probe is an ES cell.
- the term "stringent conditions" refers to conditions under which a DNA having a homology of about 70% or more, preferably about 80% or more, particularly preferably about 90% or more in a nucleotide sequence can hybridize. The stringency can be adjusted by appropriately changing the temperature, salt concentration, and the like during the hybridization reaction or washing. More preferred conditions are conditions under which DNA having a homology of 95% or more can hybridize.
- the ECAT2 gene is reported as a gene whose expression decreases when EC cells are stimulated with retinoic acid, pH34 (D ferentiation 46: 61-67, 1991), and according to the database of RIKEN (ES cell specific gene) 1.
- the ECAT3 gene is a gene encoding a mouse protein having an F box, and its expression has been reported to be observed in testis and ovaries (Current Biology 9: 1180-1182, 1999).
- the ECAT7 gene has been reported as DNMT3, a protein similar to DNMT3 that performs DNA methylation (Genomics 65: 293-298, 2000).
- the ECAT 9 gene is a growth factor called GDF3 in mice, Jones CM et al., Mol Endocrinol.
- the term “gene having extremely high homology” specifically means a gene that hybridizes with the ECAT 5 gene under stringent conditions, and if this requirement is satisfied.
- the base sequence of the ECAT5 gene (SEQ ID NO: 17 in the Sequence Listing) one or several bases may be deleted, substituted or added. Specifically, it is a gene having about 70% or more, preferably about 80% or more, more preferably about 90% or more, particularly preferably 95% or more homology with the ECAT5 gene.
- “a protein that behaves similarly” means a protein having the characteristics of the ECAT 5 protein that is specifically expressed in ES cells. In the sequence listing SEQ ID NO: 18), one or several amino acids may be deleted, substituted or added.
- the ES cell selection probe of the present invention includes various ECAT genes and hECA T gene sequences, which are constructed based on the sequences of SEQ ID NOS: 9, 11, 13, 15, 15, 17, 19, 21, 23 or 41, and a DNA fragment comprising a continuous partial sequence of 20 or more bases which does not include a repeated sequence in the base sequence described in SEQ ID NO: 27, 29, 31, 33, 35, 37 or 43 is included.
- the DNA fragment is not particularly limited as long as it can hybridize with the ECAT gene or the hECAT gene.
- Preferred examples thereof include the NA fragments described in SEQ ID NOs: 1 to 8 in the sequence listing.
- ECAT2 ECAT3, ECAT4, ECAT5, ECAT7, ECAT8 and ECAT9
- human ECAT gene was present (described below: hECAT 2, hECAT 3, hEC AT 4, hECAT 5, h ECAT 7, and hECAT 8, and hEC AT 9 genes, respectively).
- the nucleotide sequence of the CAT 8 gene and the protein encoded by the nucleotide sequence The quality amino acid sequence has not yet been determined. Accordingly, the present invention provides hECAT3, hECAT5, and hECAT8 genes and proteins, genes having extremely high homology to them, and proteins exhibiting similar behavior.
- the term “genes with extremely high homology” and “proteins exhibiting similar behavior” specifically refer to those that hybridize with the hECAT3, hECAT5 or hECAT8 gene under stringent conditions. Means the gene, and if this requirement is satisfied, one or several of the nucleotide sequences of the hECAT3, hECAT5 or hECAT8 gene (SEQ ID NO: 29, SEQ ID NO: 33, SEQ ID NO: 37, respectively) Bases may be deleted, substituted or added. Specifically, they are genes having about 70% or more, preferably about 80% or more, more preferably about 90% or more, particularly preferably 95% or more homology with those genes.
- a protein having the same behavior means a protein having the characteristics of the hECAT3 protein, hECAT5 protein or hECAT8 protein.
- One or several amino acids may be deleted, substituted or added in the acid sequence (SEQ ID NO: 30; SEQ ID NO: 34; SEQ ID NO: 38).
- the probe of the present invention can be prepared by a technique known in the art.
- this probe uses DNA isolated from EST of the corresponding ECAT gene by digestion with restriction enzymes, and complementary DNA (cDNA) prepared from genomic DNA or mRNA derived from ES cells as a ⁇ -type PCR. It can be prepared as DNA obtained by amplification by a method, DNA synthesized chemically, or DNA constructed by appropriately combining these methods.
- the present invention provides a method for screening ES cells, which comprises analyzing the expression status of a gene that is specifically expressed in ES cells.
- genes specifically expressed in ES cells is synonymous with the above-mentioned ECAT gene or hECAT gene, and specifically, ECAT1, ECAT2, and ECAT3 genes. , ECA
- T4 gene ECAT5 gene, ECAT6 gene, ECAT7 gene, ECAT 8 genes, ECAT9 gene, hECAT2 gene, hECAT3 gene, hECAT4 gene, hECAT5 gene, hECAT7 gene, hECAT8 gene and hECAT9 gene, and Oct-3 / 4 gene HOct-3 / 4 gene.
- ES cell screening is carried out by analyzing the expression status of ES cell-specific expressed genes or ES cell-specific expressed proteins encoded by them.
- the above-described probe for ES cell selection can be used.
- a probe comprising a polynucleotide containing DNA encoding Oct-3Z4 in combination.
- Such a probe may be labeled with a fluorescent substance, an enzyme, a radioisotope, or the like.
- the antibody is not particularly limited as long as it can specifically bind to the protein, and may be any of a polyclonal antibody, a monoclonal antibody, and a functional fragment thereof. These antibodies or fragments thereof may be labeled with a fluorescent substance, an enzyme, a radioisotope, or the like.
- these may be commercially available ones, and can be appropriately prepared according to a conventional method.
- the present invention also relates to an expression vector containing the ES cell-specific expression gene or the gene encoding the ES cell-specific expression protein.
- the ES cell-specific expressed gene is as described above, and the gene coding for the ES cell-specific expressed protein is specifically a gene encoding ECAT 1 (SEQ ID NO: 10). , ECAT2 (SEQ ID NO: 12) or hECAT 2 (SEQ ID NO: 2)
- ECAT4 (SEQ ID NO: 16), which encodes (SEQ ID NO: 30) Or a gene encoding hECAT4 (SEQ ID NO: 32), a gene encoding ECAT 5 (SEQ ID NO: 18) or hECAT5 (SEQ ID NO: 34), ECAT6 (SEQ ID NO: 20) Genes encoding ECAT7 (SEQ ID NO: 22) or hECAT7 (SEQ ID NO: 36), genes encoding ECAT8 (SEQ ID NO: 24) or hECAT8 (SEQ ID NO: 38) and E CAT 9 (SEQ ID NO: 42) or h E CAT 9
- Such an expression vector preferably has a function of suppressing differentiation by being expressed in a cell, particularly an ES cell, due to the nature of the gene contained therein. That is, it is a vector for forcibly expressing a differentiation-suppressing gene (hereinafter, also referred to as a differentiation-forcing gene forced expression vector).
- the expression vector of the present invention can maintain the replication or autonomously propagate in various animal cells,
- the expression vector can be prepared based on a conventional genetic engineering technique, for example, according to a basic book such as Molecular Cloning 2nd Ed., Cold Spring Harbor Laboratory Press (198 9 ).
- it is a viral vector, for example, a DNA or RNA virus such as a retrovirus, an adenovirus, an adeno-associated virus, a henorevirus, a vaccinia virus / less, a box inoles, a polio inoles, or a sylbis virus.
- a DNA or RNA virus such as a retrovirus, an adenovirus, an adeno-associated virus, a henorevirus, a vaccinia virus / less, a box inoles, a polio inoles, or a sylbis virus.
- It is prepared by incorporating ES cell-specific expressed genes into the cell.
- it is also possible to introduce a desired promoter region, drug resistance gene region, and expression control region
- Introduction of the expression vector of the present invention into cells is carried out by a conventionally known method such as transfection, ribophenetion, microinjection, impact missile, and electorifice poration.
- the produced expression vector of the present invention is integrated and expressed in a host cell is determined by, for example, determining the amount of the protein (polypeptide) produced by the expression of the introduced ECAT gene, for example, EL. It can be confirmed by measuring by the ISA method or the like. Further, the ECAT gene can be used not only as a probe for determining whether or not the cell is an ES cell, but also for selectively separating the ES cell from a mixture of the ES cell and other types of cells. The present inventors have prepared a targeting vector for knocking in a drug selection gene into the protein translation region of each ECAT gene, and have established ES cells that have undergone homologous recombination using the vector.
- the method was carried out according to the method described in Japanese Patent Publication No. 9-500000 (corresponding to U.S. Pat.
- ECAT4 gene or ECAT5 gene were cultured in the presence of G418, no differentiated cells were found in the selected cells.
- EST is a method in which a large number of cDNA clones are randomly extracted from cDNA libraries derived from various cells and organs, and the sequence of the 5 'or 3' stump is analyzed for only one reaction and registered in the public database. It is. EST can be said to be a catalog of genes expressed in each cell and organ. More than 1 million clones have been registered from mice and more than 30,000 clones have been registered only from mouse ES cells.
- Each set consists of at least one EST or known gene.
- Digital differential display method was used as an analysis method. This method calculates the frequency of each set in a library of specified cells or organs, that is, the number of EST clones contained in each set divided by the total number of EST entries from that library, and calculates This is a method for examining the frequency of expression between organs.
- Gene expression frequency analysis was performed on a group of libraries derived from the following five cells and organs by the digital differential display method. The number in parentheses in each group indicates the number of clones analyzed. All corresponding libraries from Groups 1 to 5 were analyzed. Since the data amount of the six groups was enormous, 23 libraries were extracted to include as many organs and cells as possible.
- ESII was present only in cell-derived libraries.
- the URL of the database and horn analysis program used are as follows.
- nttp // www. ncbi. nlm. nin. gov / UniGene / Mm. Home, html
- Oct_3 / 4 DNA fragment containing the sequence described in SEQ ID NO: 25 prepared by cutting out plasmid C1 in BS KS (Cell 60: 461-472, 1990) with EcoRI.
- ECAT 1 a DNA fragment containing the sequence described in SEQ ID NO: 1 prepared by cutting out Mm. 31054EST (#AI 467128) with SalI / NotI.
- ECAT2 DNA fragment containing the sequence described in SEQ ID NO: 2 prepared by cutting out pH34EST (# AA473473) with SalI / NotI. .
- ECAT 3 DNA fragment containing the sequence described in SEQ ID NO: 3 prepared by cutting out FBX15EST (# AA571680) with SalI / NotI.
- ECAT 4 DNA fragment containing the sequence shown in SEQ ID NO: 4 prepared by cutting out the home box coding region by PCR and excising it from a TA-cloned plasmid for use as a gateway with EcoRI.
- ECAT 5 a DNA fragment containing the sequence described in SEQ ID NO: 5 prepared by cutting out a plasmid obtained by RT-PCR using ER as S118 / RACE11 and excising it from TA-cloned plasmid with EcoRI.
- ECAT6 DNA fragment containing the sequence shown in SEQ ID NO: 6 which is a keratin-E PCR product (48927 SZ48927AS).
- ECAT7 a DNA fragment containing the sequence described in SEQ ID NO: 7 prepared by cutting out from the DNMT3 LEST clone (AA 895770, pBSSK-dnmt31) with EcoRI / XhoI.
- ECAT 8 MNA. 77010 A DNA fragment containing the sequence described in SEQ ID NO: 8 prepared by cutting out the ECE product from the TA C cloning plasmid with EcoRI.
- Labeling of the probe with 32 P-d CTP was performed using Amersham Pharmacia's megalabeling kit. Hybridization was performed using Funakoshi Quickhyb. The signonole analysis after washing was performed using BAS5000 of Fuji Film.
- ECAT4 gene As a result of attempting to identify the full-length cDNA by the RACE method, translation regions were identified for the ECAT4 gene, ECAT5 gene, and ECAT6 gene.
- ECAT4 gene As a result of searching the predicted amino acid sequence using B1astP, it was found that the ECAT4 gene had a homeobox, the ECAT5 gene had homology to the oncogene H-Ras, The CAT 6 gene was found to be similar to keratin, and the ECAT 9 gene, which was newly confirmed to be an ECAT gene, was found to be a growth factor called GDF-3.
- the CAT 5 gene was not included in the database.
- the ECAT2 gene is listed as ES cell specific gene (ESG) 1 in the RIKEN database, but there was no information on the expression of the other eight genes in ES cells.
- ECAT 9 As for the gene, the hECAT9 gene exists as hGDF3 (Caricasole et al., Supra). No human orthologs could be identified for the EC AT1 and EC AT6 genes.
- RNA from 13 types of human adult organs purchased from Saturation Technology or Funakoshi
- total RNA from human mesenchymal stem cells purchased from Takara
- monkey ES cells not available
- Total RNA derived from differentiation and induced by retinoic acid provided by Prof. Nakatsuji at the Institute of Regenerative Medicine at Kyoto University, was analyzed by Northern blotting.
- hECAT2, 4, 7, 8, 9 and h O ct 3/4 the total length c DNA of the corresponding EST clones in was used as a probe.
- Hybridization was carried out in the same manner as in the analysis of mouse ECAT in Example 1 above, but the reaction and washing temperature were set low (50 ° C) so that monkey RNA could be detected with a human probe.
- ECAT genes that are specifically expressed in mouse ES cells can be provided.
- seven corresponding human ECAT genes are provided.
- selective cell markers for ES cells can be obtained.
- drug resistance In the selection of ES cells by combining them with genes, trials of inducing ES cell-like cells from somatic cells, etc., it is more effective than when the Oct_3 / 4 gene or its fragment is used alone, and regeneration It is considered useful in medical and other settings.
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Developmental Biology & Embryology (AREA)
- Gynecology & Obstetrics (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- Reproductive Health (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Description
Claims
Priority Applications (8)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ES02733265.9T ES2544854T3 (es) | 2001-05-31 | 2002-05-31 | Genes con expresión específica de células ES |
EP02733265.9A EP1403366B1 (en) | 2001-05-31 | 2002-05-31 | Genes with es cell-specific expression |
AU2002306374A AU2002306374B2 (en) | 2001-05-31 | 2002-05-31 | Genes with ES cell-specific expression |
JP2003500255A JP4183614B2 (ja) | 2001-05-31 | 2002-05-31 | Es細胞特異的発現遺伝子 |
US10/479,334 US7250255B2 (en) | 2001-05-31 | 2002-05-31 | Genes with ES cell-specific expression |
US11/483,880 US8158766B2 (en) | 2001-05-31 | 2006-07-10 | Genes with ES cell-specific expression |
AU2008201280A AU2008201280B2 (en) | 2001-05-31 | 2008-03-19 | Genes with ES cell-specific expression |
US13/423,734 US8597895B2 (en) | 2001-05-31 | 2012-03-19 | Genes with ES cell-specific expression |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2001165927 | 2001-05-31 | ||
JP2001-165927 | 2001-05-31 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US10479334 A-371-Of-International | 2002-05-31 | ||
US11/483,880 Division US8158766B2 (en) | 2001-05-31 | 2006-07-10 | Genes with ES cell-specific expression |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2002097090A1 true WO2002097090A1 (en) | 2002-12-05 |
Family
ID=19008539
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2002/005350 WO2002097090A1 (en) | 2001-05-31 | 2002-05-31 | Genes with es cell-specific expression |
Country Status (6)
Country | Link |
---|---|
US (3) | US7250255B2 (ja) |
EP (2) | EP2354227B1 (ja) |
JP (2) | JP4183614B2 (ja) |
AU (2) | AU2002306374B2 (ja) |
ES (2) | ES2633306T3 (ja) |
WO (1) | WO2002097090A1 (ja) |
Cited By (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004072226A2 (en) * | 2003-02-13 | 2004-08-26 | Reprocell Inc. | Marker for undifferentiated state of cell and composition and method for separation and preparation of stem cells |
WO2005080598A1 (ja) * | 2004-02-19 | 2005-09-01 | Dainippon Sumitomo Pharma Co., Ltd. | 体細胞核初期化物質のスクリーニング方法 |
WO2006035741A1 (ja) * | 2004-09-29 | 2006-04-06 | Dainippon Sumitomo Pharma Co., Ltd. | Es細胞特異的発現遺伝子及びその利用 |
WO2007069666A1 (ja) * | 2005-12-13 | 2007-06-21 | Kyoto University | 核初期化因子 |
JP2008536803A (ja) * | 2005-02-23 | 2008-09-11 | ユニバーシティ オブ バージニア パテント ファウンデーション | 精子タンパク質に対する新規卵受容体 |
US8071369B2 (en) | 2003-11-26 | 2011-12-06 | Whitehead Institute For Biomedical Research | Compositions for reprogramming somatic cells |
US8129187B2 (en) | 2005-12-13 | 2012-03-06 | Kyoto University | Somatic cell reprogramming by retroviral vectors encoding Oct3/4. Klf4, c-Myc and Sox2 |
US8211697B2 (en) | 2007-06-15 | 2012-07-03 | Kyoto University | Induced pluripotent stem cells produced using reprogramming factors and a rho kinase inhibitor or a histone deacetylase inhibitor |
US8278104B2 (en) | 2005-12-13 | 2012-10-02 | Kyoto University | Induced pluripotent stem cells produced with Oct3/4, Klf4 and Sox2 |
US8791248B2 (en) | 2007-12-10 | 2014-07-29 | Kyoto University | Nuclear reprogramming factor comprising miRNA and a protein factor |
US9213999B2 (en) | 2007-06-15 | 2015-12-15 | Kyoto University | Providing iPSCs to a customer |
US9382515B2 (en) | 2007-04-07 | 2016-07-05 | Whitehead Institute For Biomedical Research | Reprogramming of somatic cells |
US9497943B2 (en) | 2008-06-13 | 2016-11-22 | Whitehead Institute For Biomedical Research | Nucleic acid constructs encoding reprogramming factors linked by self-cleaving peptides |
US9499797B2 (en) | 2008-05-02 | 2016-11-22 | Kyoto University | Method of making induced pluripotent stem cells |
US9683232B2 (en) | 2007-12-10 | 2017-06-20 | Kyoto University | Efficient method for nuclear reprogramming |
US9803012B2 (en) | 2005-02-23 | 2017-10-31 | University Of Virginia Patent Foundation | Compositions and methods for regulating SAS1R |
JP2018126171A (ja) * | 2009-11-12 | 2018-08-16 | ブイビーアイ テクノロジーズ, エルエルシー | 胞子様細胞の亜集団およびその用途 |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090203141A1 (en) * | 2003-05-15 | 2009-08-13 | Shi-Lung Lin | Generation of tumor-free embryonic stem-like pluripotent cells using inducible recombinant RNA agents |
US20090227032A1 (en) * | 2005-12-13 | 2009-09-10 | Kyoto University | Nuclear reprogramming factor and induced pluripotent stem cells |
US20090239217A1 (en) * | 2007-10-08 | 2009-09-24 | University Of Florida Research Foundation, Inc. | Stem-like cells in bone sarcomas |
US20110033931A1 (en) * | 2007-11-29 | 2011-02-10 | Children's Hospital Of Orange County | De-differentiation of human cells |
CA2621155A1 (en) * | 2008-02-29 | 2009-08-29 | James Ellis | Stem cell expression cassettes |
US9394538B2 (en) | 2008-05-07 | 2016-07-19 | Shi-Lung Lin | Development of universal cancer drugs and vaccines |
AU2017270234B2 (en) | 2016-05-25 | 2023-11-23 | Assistance Publique-Hôpitaux De Paris (Aphp) | Methods and compositions for treating cancers |
KR20200111168A (ko) | 2017-11-24 | 2020-09-28 | 인쎄름 (엥스띠뛰 나씨오날 드 라 쌍떼 에 드 라 흐쉐르슈 메디깔) | 암 치료를 위한 방법 및 조성물 |
WO2020030634A1 (en) | 2018-08-06 | 2020-02-13 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating cancers |
EP4314246A1 (en) | 2021-04-01 | 2024-02-07 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Liver organoid manufacturing methods, liver organoids obtained with the same, and uses thereof |
Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1994015965A1 (en) * | 1993-01-12 | 1994-07-21 | Johns Hopkins University School Of Medicine | Growth differentiation factor-3 |
WO1994024274A1 (en) * | 1993-04-21 | 1994-10-27 | The University Of Edinburgh | Isolation, selection and propagation of animal transgenic stem cells |
WO1999014372A1 (en) * | 1997-09-15 | 1999-03-25 | Abbott Laboratories | Reagents and methods useful for detecting diseases of the urinary tract |
WO1999018989A1 (en) * | 1997-10-16 | 1999-04-22 | Baylor College Of Medicine | F-box proteins and genes |
WO2000005248A1 (en) * | 1998-07-24 | 2000-02-03 | Eli Lilly And Company | Human gdf-3 nucleic acids, polypeptides, vectors, host cells, methods and uses thereof |
EP1074617A2 (en) * | 1999-07-29 | 2001-02-07 | Helix Research Institute | Primers for synthesising full-length cDNA and their use |
WO2001027249A1 (en) * | 1999-10-15 | 2001-04-19 | Fit Biotech Oyj Plc | Novel gene encoding a dna methyltransferase, dnmt3l |
WO2001057266A1 (en) * | 2000-02-03 | 2001-08-09 | Hyseq, Inc. | Methods and materials relating to alpha-2-macroglobulin-like polypeptides and polynucleotides |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2001009316A1 (fr) | 1999-07-29 | 2001-02-08 | Helix Research Institute | Nouveaux genes codant la proteine kinase / proteine phosphatase |
AU6180800A (en) | 1999-07-29 | 2001-02-19 | Chugai Research Institute For Molecular Medicine, Inc. | Novel gene participating in the maintenance of smooth muscle cell differentiation |
US6908748B2 (en) | 1999-07-29 | 2005-06-21 | Kenji Sobue | Genes associated with the maintenance of differentiation of smooth muscle cells |
JP2001165927A (ja) | 1999-12-08 | 2001-06-22 | Ngk Insulators Ltd | 内分泌攪乱化学物質のモニタリング方法およびその装置 |
FR2808803B1 (fr) * | 2000-05-11 | 2004-12-10 | Agronomique Inst Nat Rech | Cellules es modifiees et gene specifique de cellules es |
-
2002
- 2002-05-31 ES ES10010750.7T patent/ES2633306T3/es not_active Expired - Lifetime
- 2002-05-31 WO PCT/JP2002/005350 patent/WO2002097090A1/ja active Application Filing
- 2002-05-31 EP EP10010750.7A patent/EP2354227B1/en not_active Expired - Lifetime
- 2002-05-31 JP JP2003500255A patent/JP4183614B2/ja not_active Expired - Lifetime
- 2002-05-31 AU AU2002306374A patent/AU2002306374B2/en not_active Ceased
- 2002-05-31 US US10/479,334 patent/US7250255B2/en not_active Expired - Lifetime
- 2002-05-31 EP EP02733265.9A patent/EP1403366B1/en not_active Expired - Lifetime
- 2002-05-31 ES ES02733265.9T patent/ES2544854T3/es not_active Expired - Lifetime
-
2006
- 2006-07-10 US US11/483,880 patent/US8158766B2/en not_active Expired - Fee Related
-
2008
- 2008-03-19 AU AU2008201280A patent/AU2008201280B2/en not_active Ceased
- 2008-06-06 JP JP2008149839A patent/JP4829272B2/ja not_active Expired - Lifetime
-
2012
- 2012-03-19 US US13/423,734 patent/US8597895B2/en not_active Expired - Lifetime
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1994015965A1 (en) * | 1993-01-12 | 1994-07-21 | Johns Hopkins University School Of Medicine | Growth differentiation factor-3 |
WO1994024274A1 (en) * | 1993-04-21 | 1994-10-27 | The University Of Edinburgh | Isolation, selection and propagation of animal transgenic stem cells |
WO1999014372A1 (en) * | 1997-09-15 | 1999-03-25 | Abbott Laboratories | Reagents and methods useful for detecting diseases of the urinary tract |
WO1999018989A1 (en) * | 1997-10-16 | 1999-04-22 | Baylor College Of Medicine | F-box proteins and genes |
WO2000005248A1 (en) * | 1998-07-24 | 2000-02-03 | Eli Lilly And Company | Human gdf-3 nucleic acids, polypeptides, vectors, host cells, methods and uses thereof |
EP1074617A2 (en) * | 1999-07-29 | 2001-02-07 | Helix Research Institute | Primers for synthesising full-length cDNA and their use |
WO2001027249A1 (en) * | 1999-10-15 | 2001-04-19 | Fit Biotech Oyj Plc | Novel gene encoding a dna methyltransferase, dnmt3l |
WO2001057266A1 (en) * | 2000-02-03 | 2001-08-09 | Hyseq, Inc. | Methods and materials relating to alpha-2-macroglobulin-like polypeptides and polynucleotides |
Non-Patent Citations (4)
Title |
---|
AAPOLA U. ET AL.: "Isolation and initial characterization of a novel zinc finger gene, DNMT3L, on 21q22.3, related to the cytosine-5-methyltransferase 3 gene family", GENOMICS, vol. 65, no. 3, May 2000 (2000-05-01), pages 293 - 298, XP004439370 * |
See also references of EP1403366A4 * |
TAKEDA J., SEIO S., BELL G.I.: "Human Oct3 gene family: cDNA sequences, alternative splicing, gene organization, chromosomal location and expression at low levels in adult tissues", NUCLEIC ACIDS RESEARCH, vol. 20, no. 17, September 1992 (1992-09-01), pages 4613 - 4620, XP002995407 * |
WINSTON J.T. ET AL.: "A family of mammalian F-box proteins", CURRENT BIOLOGY, vol. 9, no. 20, October 1999 (1999-10-01), pages 1180 - 1182, XP000960309 * |
Cited By (45)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2006521792A (ja) * | 2003-02-13 | 2006-09-28 | 国立大学法人京都大学 | 細胞の未分化状態マーカーならびに幹細胞の分離および調製のための組成物および方法 |
WO2004072226A2 (en) * | 2003-02-13 | 2004-08-26 | Reprocell Inc. | Marker for undifferentiated state of cell and composition and method for separation and preparation of stem cells |
WO2004072226A3 (en) * | 2003-02-13 | 2006-01-19 | Reprocell Inc | Marker for undifferentiated state of cell and composition and method for separation and preparation of stem cells |
US11655459B2 (en) | 2003-11-26 | 2023-05-23 | Whitehead Institute For Biomedical Research | Methods for reprogramming somatic cells |
US10017744B2 (en) | 2003-11-26 | 2018-07-10 | Whitehead Institute For Biomedical Research | Methods for reprogramming somatic cells |
US11987815B2 (en) | 2003-11-26 | 2024-05-21 | Whitehead Institute For Biomedical Research | Methods for reprogramming somatic cells |
US8927279B2 (en) | 2003-11-26 | 2015-01-06 | Whitehead Institute For Biomedical Research | Methods for reprogramming somatic cells |
US8932856B2 (en) | 2003-11-26 | 2015-01-13 | Whitehead Institute For Biomedical Research | Methods for reprogramming somatic cells |
US8940536B2 (en) | 2003-11-26 | 2015-01-27 | Whitehead Institute For Biomedical Research | Methods for making somatic cells more susceptible to reprogramming |
US8951797B2 (en) | 2003-11-26 | 2015-02-10 | Whitehead Institute For Biomedical Research | Compositions for identifying reprogramming factors |
US9670464B2 (en) | 2003-11-26 | 2017-06-06 | Whitehead Institute For Biomedical Research | Methods for reprogramming somatic cells |
US8071369B2 (en) | 2003-11-26 | 2011-12-06 | Whitehead Institute For Biomedical Research | Compositions for reprogramming somatic cells |
US10457917B2 (en) | 2003-11-26 | 2019-10-29 | Whitehead Institute For Biomedical Research | Methods for reprogramming somatic cells |
JP4901471B2 (ja) * | 2004-02-19 | 2012-03-21 | 国立大学法人京都大学 | 体細胞核初期化物質のスクリーニング方法 |
US7964401B2 (en) | 2004-02-19 | 2011-06-21 | Kyoto University | Screening method for somatic cell nuclear reprogramming substance affecting ECAT2 and ECAT3 |
WO2005080598A1 (ja) * | 2004-02-19 | 2005-09-01 | Dainippon Sumitomo Pharma Co., Ltd. | 体細胞核初期化物質のスクリーニング方法 |
JPWO2005080598A1 (ja) * | 2004-02-19 | 2007-08-30 | 伸弥 山中 | 体細胞核初期化物質のスクリーニング方法 |
WO2006035741A1 (ja) * | 2004-09-29 | 2006-04-06 | Dainippon Sumitomo Pharma Co., Ltd. | Es細胞特異的発現遺伝子及びその利用 |
US7803920B2 (en) | 2004-09-29 | 2010-09-28 | Shinya Yamanaka | ECAT16 gene expressed specifically in ES cells and utilization of the same |
JPWO2006035741A1 (ja) * | 2004-09-29 | 2008-05-15 | 伸弥 山中 | Es細胞特異的発現遺伝子及びその利用 |
US9803012B2 (en) | 2005-02-23 | 2017-10-31 | University Of Virginia Patent Foundation | Compositions and methods for regulating SAS1R |
JP2008536803A (ja) * | 2005-02-23 | 2008-09-11 | ユニバーシティ オブ バージニア パテント ファウンデーション | 精子タンパク質に対する新規卵受容体 |
CN103773804A (zh) * | 2005-12-13 | 2014-05-07 | 国立大学法人京都大学 | 核重新编程因子 |
JP2011188860A (ja) * | 2005-12-13 | 2011-09-29 | Kyoto Univ | 誘導多能性幹細胞 |
JP2014000083A (ja) * | 2005-12-13 | 2014-01-09 | Kyoto Univ | 誘導多能性幹細胞 |
EA018039B1 (ru) * | 2005-12-13 | 2013-05-30 | Киото Юниверсити | Ядерный фактор перепрограммирования |
US8278104B2 (en) | 2005-12-13 | 2012-10-02 | Kyoto University | Induced pluripotent stem cells produced with Oct3/4, Klf4 and Sox2 |
EP3418297A1 (en) * | 2005-12-13 | 2018-12-26 | Kyoto University | Nuclear reprogramming factor |
US8048999B2 (en) | 2005-12-13 | 2011-11-01 | Kyoto University | Nuclear reprogramming factor |
US8129187B2 (en) | 2005-12-13 | 2012-03-06 | Kyoto University | Somatic cell reprogramming by retroviral vectors encoding Oct3/4. Klf4, c-Myc and Sox2 |
WO2007069666A1 (ja) * | 2005-12-13 | 2007-06-21 | Kyoto University | 核初期化因子 |
EA014166B1 (ru) * | 2005-12-13 | 2010-10-29 | Киото Юниверсити | Ядерный фактор перепрограммирования |
US9714414B2 (en) | 2007-04-07 | 2017-07-25 | Whitehead Institute For Biomedical Research | Reprogramming of somatic cells |
US10093904B2 (en) | 2007-04-07 | 2018-10-09 | Whitehead Institute For Biomedical Research | Reprogramming of somatic cells |
US9382515B2 (en) | 2007-04-07 | 2016-07-05 | Whitehead Institute For Biomedical Research | Reprogramming of somatic cells |
US8211697B2 (en) | 2007-06-15 | 2012-07-03 | Kyoto University | Induced pluripotent stem cells produced using reprogramming factors and a rho kinase inhibitor or a histone deacetylase inhibitor |
US9714433B2 (en) | 2007-06-15 | 2017-07-25 | Kyoto University | Human pluripotent stem cells induced from undifferentiated stem cells derived from a human postnatal tissue |
US9213999B2 (en) | 2007-06-15 | 2015-12-15 | Kyoto University | Providing iPSCs to a customer |
US8257941B2 (en) | 2007-06-15 | 2012-09-04 | Kyoto University | Methods and platforms for drug discovery using induced pluripotent stem cells |
US9683232B2 (en) | 2007-12-10 | 2017-06-20 | Kyoto University | Efficient method for nuclear reprogramming |
US8791248B2 (en) | 2007-12-10 | 2014-07-29 | Kyoto University | Nuclear reprogramming factor comprising miRNA and a protein factor |
US9499797B2 (en) | 2008-05-02 | 2016-11-22 | Kyoto University | Method of making induced pluripotent stem cells |
US11851670B2 (en) | 2008-06-13 | 2023-12-26 | Whitehead Institute For Biomedical Research | Nucleic acid constructs encoding reprogramming factors linked by self-cleaving peptides |
US9497943B2 (en) | 2008-06-13 | 2016-11-22 | Whitehead Institute For Biomedical Research | Nucleic acid constructs encoding reprogramming factors linked by self-cleaving peptides |
JP2018126171A (ja) * | 2009-11-12 | 2018-08-16 | ブイビーアイ テクノロジーズ, エルエルシー | 胞子様細胞の亜集団およびその用途 |
Also Published As
Publication number | Publication date |
---|---|
ES2544854T3 (es) | 2015-09-04 |
AU2008201280A1 (en) | 2008-04-17 |
US20120178911A1 (en) | 2012-07-12 |
US20040137460A1 (en) | 2004-07-15 |
AU2002306374B2 (en) | 2008-01-24 |
JP2009000108A (ja) | 2009-01-08 |
JP4829272B2 (ja) | 2011-12-07 |
EP1403366A1 (en) | 2004-03-31 |
AU2008201280B2 (en) | 2010-06-03 |
US8158766B2 (en) | 2012-04-17 |
US20060292620A1 (en) | 2006-12-28 |
JP4183614B2 (ja) | 2008-11-19 |
ES2633306T3 (es) | 2017-09-20 |
EP1403366A4 (en) | 2006-02-01 |
US7250255B2 (en) | 2007-07-31 |
EP2354227A1 (en) | 2011-08-10 |
JPWO2002097090A1 (ja) | 2004-09-09 |
EP2354227B1 (en) | 2017-04-19 |
EP1403366B1 (en) | 2015-05-27 |
US8597895B2 (en) | 2013-12-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP4829272B2 (ja) | Es細胞特異的発現遺伝子 | |
US5652128A (en) | Method for producing tagged genes, transcripts, and proteins | |
Kazmierczak et al. | Description of a novel fusion transcript between HMGI-C, a gene encoding for a member of the high mobility group proteins, and the mitochondrial aldehyde dehydrogenase gene | |
Beard et al. | High-throughput genomic mapping of vector integration sites in gene therapy studies | |
JPH07132088A (ja) | ウシ胚の性の識別方法 | |
CN109837273A (zh) | 一种crispr辅助dna靶向富集方法及其应用 | |
CN109402115B (zh) | 靶向Rett突变基因RNA的gRNA及Rett突变基因的检测方法、检测试剂盒 | |
JP2002506635A (ja) | 遺伝子の同定に対する選択サブトラクションアプローチ | |
US20060228714A1 (en) | Nucleic acid representations utilizing type IIB restriction endonuclease cleavage products | |
JP3844656B2 (ja) | 動物細胞の形質転換のための方法 | |
CN111132671A (zh) | 治疗印记缺陷相关疾病的组合物和方法 | |
Kohwi-Shigematsu et al. | Identification of base-unpairing region-binding proteins and characterization of their in vivo binding sequences | |
CN112899238A (zh) | 基于RNA-m6A修饰水平的化合物筛选细胞模型及其构建与应用 | |
JP2006517181A (ja) | Bbb−特異的タンパク質およびその断片を同定する方法 | |
Ligon et al. | PCOLCE deletion and expression analyses in uterine leiomyomata | |
Kim et al. | A subtractive PCR-based cDNA library made from fetal thymic stromal cells | |
JP2001505417A (ja) | 生物の成長に必須の遺伝子の同定方法 | |
Miura et al. | Transfer of spermatogenesis‐related cDNAs into eel testis germ‐somatic cell coculture pellets by electroporation: Methods for analysis of gene function | |
CN116376975B (zh) | 激活异染色质基因的方法及应用 | |
JP2009045004A (ja) | クロマチン制御因子による幹細胞未分化制御方法 | |
WO2015129655A1 (ja) | Dnajb1-prkaca遺伝子の検出方法 | |
JP4995654B2 (ja) | クロマチンタンパク質又はその変異体による幹細胞未分化制御方法。 | |
JP2003061666A (ja) | 多段階差次的クローニング技術と細胞増殖制御遺伝子 | |
JP2006262726A (ja) | マガキの炭酸脱水酵素遺伝子 | |
Hertzke | pSV3neo transfected bovine fetal fibroblast cell lines |
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 OM PH PL PT RO RU SD SE SG SI SK SL TJ TM TN TR TT TZ UA UG US UZ VN YU ZA ZM 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 ZM 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 |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2003500255 Country of ref document: JP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 10479334 Country of ref document: US |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2002306374 Country of ref document: AU |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2002733265 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 2002733265 Country of ref document: EP |
|
REG | Reference to national code |
Ref country code: DE Ref legal event code: 8642 |
|
ENP | Entry into the national phase |
Ref document number: 2002306374 Country of ref document: AU Date of ref document: 20020531 Kind code of ref document: B |