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METHOD FOR SITE-SPECIFIC INSERTION OF FOREIGN DNA INTO ANIMAL CELL GENOME AND CELL OBTAINED USING SAME

外国特許コード F170009072
整理番号 (S2015-1830-N0)
掲載日 2017年5月29日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2016JP059174
国際公開番号 WO 2017029833
国際出願日 平成28年3月23日(2016.3.23)
国際公開日 平成29年2月23日(2017.2.23)
優先権データ
  • 特願2015-162612 (2015.8.20) JP
  • 2015US-62265425 (2015.12.10) US
発明の名称 (英語) METHOD FOR SITE-SPECIFIC INSERTION OF FOREIGN DNA INTO ANIMAL CELL GENOME AND CELL OBTAINED USING SAME
発明の概要(英語) Provided is a method for site-specific insertion of a foreign DNA into an animal cell genome, said method comprising: (1) a step for constructing a donor plasmid in which 100-300 bp DNAs having a sequence homologous to an insertion site on a genome are ligated to the upstream and downstream of a 0.1-10 kbp foreign DNA; and (2) a step for inserting the 0.1-10 kbp foreign DNA into a target site in the genome by genome editing through homologous recombination repair with the use of the donor plasmid obtained above.
特許請求の範囲(英語) [claim1]
1. The animal cell genome region unique imported DNA insertion method of possessing the following process.
High-level and downstream of the coming from abroad DNA of (1)0.1kbp-10kbp, the process which draws up the donor plasmid which connects DNA of 100bp-300bp which possesses the region and the homologous arrangement which insert on the genome. (2) the process which inserts the coming from abroad DNA of 0.1kbp-10kbp in the region which is made purpose of the genome with the genome compilation with the homologous rearrangement restoration which uses the donor plasmid which is obtained.
[claim2]
2. The animal cell, to stocks the human origin cell which is converted, to stocks the mouse origin cell which is converted, to stocks in the claim 1 which is the cell which is chosen from the chicken origin cell, the human ES cell, the mouse ES cell, the human iPS cell and the mouse iPS cell which are converted genome region unique imported DNA insertion method of statement.
[claim3]
3. The animal cell, in the claim 1 which is the cell which is chosen from the human HCT116 cell, the human HT1080 cell, the human NALM6 cell, the chicken DT40 cell, the human ES cell, the human iPS cell, the mouse ES cell and the mouse iPS cell or 2 genome region unique imported DNA insertion method of statement.
[claim4]
4. The genome editing due to homologous rearrangement restoration, either of the claim 1-3 which is the imported DNA insertion method which utilizes the homologous rearrangement restoration which is chosen from the CRISPR-Cas9 system, the TALEN system or the Zn finger nuclease system in one section genome region unique imported DNA insertion method of statement.
[claim5]
5. The genome editing due to homologous rearrangement restoration, either of the claim 1-4 which is the imported DNA insertion method which utilizes the homologous rearrangement restoration which is induced by the CRISPR-Cas9 system in one section genome region unique imported DNA insertion method of statement.
[claim6]
6. Homology arrangement DNA of the aforementioned insertion genome region, either of the claim 1-5 which is something which is obtained by PCR or DNA synthesis in one section genome region unique imported DNA insertion method of statement.
[claim7]
7. Coming from abroad DNA, tag addition coming from abroad DNA, either of the claim 1-6 which is the coming from abroad DNA which is chosen from promoter arrangement, copying conclusion arrangement, functional gene arrangement, the medicine selected marker gene and those combinations in one section genome region unique imported DNA insertion method of statement.
[claim8]
8. In safe harbor sitting position transport inhibiter response 1 (TIR1) the cord/code the cell which features that it possesses the chromosome which includes the gene which is done.
[claim9]
9. The aforementioned safe harbor sitting position AAVS1 (the AAV integration site 1) in the claim 8 which is the heredity stroma the cell of statement.
[claim10]
10. Furthermore, in the claim 8 where the aforementioned chromosome operation was connected to the gene which the description above TIR1 the cord/code is done possibly, includes the inductive promoter, the virus characteristic promoter, the housekeeping gene promoter or the organizational unique promoter or 9 the cell of statement.
[claim11]
11. The aforementioned chromosome includes the inductive promoter who operation possibly is connected in the gene which the description above TIR1 the cord/code is done, in claim 10 the cell of statement.
[claim12]
12. The aforementioned inductive promoter, is selected from the group which consists of the chemical inductive promoter, the thermal shock inductive promoter, the electromagnetics inductive promoter, the nuclear receptor inductive promoter and the hormone inductive promoter, in claim 11 the cell of statement.
[claim13]
13. The aforementioned chemical inductive promoter is the tetracycline inductive promoter, in claim 12 the cell of statement.
[claim14]
14. Making use of DNA of 100-300bp which possesses the region and the homologous arrangement which are connected by the high level and downstream of coming from abroad, DNA insert on the chromosome, the cell which features that it possesses the chromosome where insertion of aforementioned coming from abroad DNA was done.
[claim15]
15. Furthermore, the aforementioned chromosome, being the high level or downstream of the aforementioned coming from abroad DNA, includes the medicine selected marker gene which is connected to the territory where it was put to DNA the description above 100-300bp, in claim 14 the cell of statement.
[claim16]
16. The first gene which the intended protein the cord/code is done and,
Mini-auxin-inducible degron which is connected to the high level or downstream of the aforementioned first gene (mAID) the cord/code the second gene which is done and,
The first chromosome which is included and,
In safe harbor sitting position the second chromosome which includes the gene which TIR1 the cord/code is done and,
The cell which features that it possesses.
[claim17]
17. The aforementioned first chromosome is connected by the high level and downstream of the aforementioned first gene and the aforementioned second gene, includes DNA of 100-300bp which possesses the region and the homologous arrangement which insertion on the chromosome does, in claim 16 the cell of statement.
[claim18]
18. The aforementioned first chromosome, being the high level or downstream of the first gene and the aforementioned second gene, includes the medicine selected marker gene which is connected to the territory where it was put to DNA the description above 100-300bp, in claim 16 or 17 the cell of statement.
[claim19]
19. Furthermore, the aforementioned chromosome operation possibly was connected to the gene which the description above TIR1 the cord/code is done, the inductive promoter, the virus characteristic promoter, the housekeeping gene promoter or the organizational unique promoter are included, either of the claim 16-18 in one section the cell of statement.
[claim20]
20. The aforementioned chromosome includes the inductive promoter who operation possibly is connected in the gene which the description above TIR1 the cord/code is done, in claim 19 the cell of statement.
[claim21]
21. The aforementioned inductive promoter, is selected from the group which consists of the chemical inductive promoter, the thermal shock inductive promoter, the electromagnetics inductive promoter, the nuclear receptor inductive promoter and the hormone inductive promoter, in claim 20 the cell of statement.
[claim22]
22. The aforementioned chemical inductive promoter is the tetracycline inductive promoter, in claim 21 the cell of statement.
[claim23]
23. Either of the claim 16-22 disassembly method of the intended protein which features that the cell of statement is used for one section.
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • INTER-UNIVERSITY RESEARCH INSTITUTE CORPORATION RESEARCH ORGANIZATION OF INFORMATION AND SYSTEMS
  • 発明者(英語)
  • KANEMAKI MASATO
  • NATSUME TOYOAKI
国際特許分類(IPC)
指定国 (WO201729833)
National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG

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