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GENETIC MODIFICATION NON-HUMAN ORGANISM, EGG CELLS, FERTILIZED EGGS, AND METHOD FOR MODIFYING TARGET GENES NEW

外国特許コード F170009118
整理番号 (AF37P001)
掲載日 2017年7月7日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2016JP085391
国際公開番号 WO 2017104404
国際出願日 平成28年11月29日(2016.11.29)
国際公開日 平成29年6月22日(2017.6.22)
優先権データ
  • 特願2015-247960 (2015.12.18) JP
発明の名称 (英語) GENETIC MODIFICATION NON-HUMAN ORGANISM, EGG CELLS, FERTILIZED EGGS, AND METHOD FOR MODIFYING TARGET GENES NEW
発明の概要(英語) The present invention provides a genetic modification non-human organism where Cas9 expression is higher and a plurality of different genes, or a plurality of sites within the same gene, can be simultaneously edited with high efficiency. This genetic modification non-human organism has at least three copies of the gene encoding Cas9 (CRISPR-associated 9) within the nuclear genome. These egg cells are derived from a genetic modification non-human organism having a nuclear genome into which at least three copies of a gene encoding CAS9 have been introduced. These fertilized eggs are obtained by allowing fertilization to occur between the egg cells and sperm derived from an organism of the same species. This method for modifying a target gene comprises a step for introducing guide RNA to cells derived from the genetic modification non-human organism, the egg cells, or the fertilized eggs.
特許請求の範囲(英語) [claim1]
1. Cas9 (CRISPR-associated 9) the cord/code the gene alteration non human living thing which features that the gene which is done at least has 3 copies in the nuclear genome.
[claim2]
2. The description above Cas9 is reforming transformation, in claim 1 the gene alteration non human living thing of statement.
[claim3]
3. The activity which cuts off one side or the both chain of the target gene to which the description above Cas9 includes target sequence it is the reforming transformation which lacks, in claim 2 the gene alteration non human living thing of statement.
[claim4]
4. Furthermore, the aforementioned nuclear genome operation possibly was connected to the gene which the description above Cas9 the cord/code is done, the CAG promoter, the revelation inductive promoter, the virus characteristic promoter or the organizational unique promoter are included, either of the claim 1-3 in one section the gene alteration non human living thing of statement.
[claim5]
5. The non human living thing is the mammal, either of the claim 1-4 in one section the gene alteration non human living thing of statement.
[claim6]
6. The gene which the description above Cas9 the cord/code is done was introduced into tandem in the aforementioned nuclear genome, either of the claim 1-5 in one section the gene alteration non human living thing of statement.
[claim7]
7. The egg cell which features that it is the gene alteration non human living thing origin which possesses the nuclear genome where the gene which Cas9 the cord/code is done at least 3 is copied is introduced.
[claim8]
8. The aforementioned egg cell does not have the nuclear genome where the gene which Cas9 the cord/code is done is introduced in the nucleus, in claim 7 the egg cell of statement.
[claim9]
9. Claim making with 7 or the egg cell of statement and the spermatozoon of similar living thing origin 8 fertilize, the fertilized egg which features that it becomes.
[claim10]
10. The aforementioned egg cell does not have the nuclear genome where the gene which Cas9 the cord/code is done is introduced in the nucleus, in claim 9 the fertilized egg of statement.
[claim11]
11. The aforementioned spermatozoon is the aforementioned similar living thing origin of wild type, claim 9 or in 10 the fertilized egg of statement.
[claim12]
12. Either of the claim 1-6 in one section the cell of gene alteration non human living thing origin of statement, in claim 7 or 8 the egg cell of statement, or either of the claim 9-11 alteration method of the target gene which features that the process which introduces guide RNA into the fertilized egg of statement in one section is had.
[claim13]
13. Either of the claim 1-6 in one section the cell of gene alteration non human living thing origin of statement, in claim 7 or 8 the egg cell of statement, or either of the claim 9-11 the process which introduces guide RNA into the fertilized egg of statement in one section and,
Cas9 which has been revealed in the cell of the aforementioned gene alteration non human living thing origin, the egg cell of the aforementioned gene alteration non human living thing origin, or inside the fertilized egg of the aforementioned gene alteration non human living thing origin PAM (Proto-spacer Adjacent Motif) the process which cuts off the aforementioned target gene with the cutting region where arrangement is located high-level and,
The process which obtains the aforementioned target gene which is altered in the territory where it is decided by the complimentary connection of the aforementioned guide RNA and the aforementioned target gene, and, having,
As for the aforementioned target gene possessing the aforementioned PAM arrangement,
As for the aforementioned guide RNA, alteration method of the target gene which features that it is something which includes the polynucleotide which consists of complimentary base arrangement in high-level base arrangement of the aforementioned PAM arrangement in the aforementioned target gene.
[claim14]
14. Either of the claim 1-6 in one section the cell of gene alteration non human living thing origin of statement, in claim 7 or 8 the egg cell of statement, or either of the claim 9-11 the process which introduces guide RNA into the fertilized egg of statement in one section and,
Cas9 which has been revealed in the cell of the aforementioned gene alteration non human living thing origin, the egg cell of the aforementioned gene alteration non human living thing origin, or inside the fertilized egg of the aforementioned gene alteration non human living thing origin 3 bases of PAM arrangement the process which cuts off the aforementioned target gene with the cutting region which is located high-level and,
The process which obtains the aforementioned target gene which is altered in the territory where it is decided by the complimentary connection of the aforementioned guide RNA and the aforementioned target gene, and, having,
As for the aforementioned target gene NGG (as for N, the optional 1 bases which are selected from the group which consists of adenine, cytosine, the thymine and guanine) possessing the PAM arrangement which consists of,
As for the aforementioned guide RNA, alteration method of the target gene which features that it is something which includes the polynucleotide where from 1 base high levels of the aforementioned PAM arrangement in the aforementioned target gene to the high level of 24 base or less of 20 bases or more consists of complimentary base arrangement in base arrangement.
[claim15]
15. In the aforementioned introduction process, when the introduction object is the cell of gene alteration non human living thing origin, guide RNA is introduced into the gene alteration non human living thing which lives, either of the claim 12-14 in one section alteration method of the target gene of statement.
[claim16]
16. In the aforementioned introduction process, when the introduction object is the fertilized egg of gene alteration non human animal origin, furthermore, the aforementioned fertilized egg is transplanted to the uterus or the uterine tube of the similar non human animal, the process which it occurs is had, either of the claim 12-14 in one section alteration method of the target gene of statement.
[claim17]
17. In the aforementioned introduction process, the aforementioned guide RNA of 2 types or more is introduced, either of the claim 12-16 in one section alteration method of the target gene of statement.
[claim18]
18. In the aforementioned introduction process, with the aforementioned guide RNA the imported gene is introduced, either of the claim 12-17 in one section alteration method of the target gene of statement.
[claim19]
19. The aforementioned imported gene does not have PAM arrangement, in claim 18 alteration method of the target gene of statement.
  • 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
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • 発明者(英語)
  • SHINDO TAKAYUKI
  • SAKURAI TAKAYUKI
国際特許分類(IPC)
指定国 (WO2017104404)
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 DJ 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 KW 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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