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1. WO2020191816 - METHOD FOR PREPARING HUMAN RETINOBLASTOMA MODEL

Publication Number WO/2020/191816
Publication Date 01.10.2020
International Application No. PCT/CN2019/081841
International Filing Date 09.04.2019
IPC
C12N 15/90 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
87Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
90Stable introduction of foreign DNA into chromosome
C12N 15/85 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79Vectors or expression systems specially adapted for eukaryotic hosts
85for animal cells
CPC
C12N 15/85
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79Vectors or expression systems specially adapted for eukaryotic hosts
85for animal cells
C12N 15/907
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
87Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
90Stable introduction of foreign DNA into chromosome
902using homologous recombination
907in mammalian cells
C12N 9/22
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes; Proenzymes; Compositions thereof
14Hydrolases (3)
16acting on ester bonds (3.1)
22Ribonucleases ; RNAses, DNAses
Applicants
  • 温州医科大学 WENZHOU MEDICAL UNIVERSITY [CN]/[CN]
Inventors
  • 金子兵 JIN, Zibing
  • 刘慧 LIU, Hui
Agents
  • 温州金瓯专利事务所(普通合伙) WENZHOU JINOU PATENT LAW OFFICE (GENERAL PARTNERSHIP)
Priority Data
201910236645.827.03.2019CN
Publication Language Chinese (ZH)
Filing Language Chinese (ZH)
Designated States
Title
(EN) METHOD FOR PREPARING HUMAN RETINOBLASTOMA MODEL
(FR) PROCÉDÉ DE PRÉPARATION D'UN MODÈLE DE RÉTINOBLASTOME HUMAIN
(ZH) 一种人视网膜母细胞瘤模型的制备方法
Abstract
(EN)
The present invention provides a method for preparing a human retinoblastoma model, comprising: (a) establishing RB1 allele mutation or knockout human pluripotent stem cell line by genetically editing the human retinoblastoma RB1 allele in human embryonic stem cells, or reprogramming the somatic cells of Rb patients carrying RB1 allele mutations to human induced pluripotent stem cells for further screening and identification; and (B) using an in vitro three-dimensional retinal differentiation system to induce RB1 allele mutation or knockout human pluripotent stem cells to differentiate into a human retinoblastoma model.
(FR)
Procédé de préparation d'un modèle de rétinoblastome humain comprenant les étapes suivantes : (A) établissement d'une mutation d'allèle RB1 ou d'une lignée de cellules souches pluripotentes humaines knock-out par modification génétique de l'allèle RB1 du rétinoblastome humain dans des cellules souches embryonnaires humaines, ou la reprogrammation des cellules somatiques de patients avec un rétinoblastome (Rb) Portant des mutations d'allèle RB1 à des cellules souches pluripotentes induites humaines pour un criblage et une identification supplémentaires; et (B) utilisation d'un système de différenciation rétinienne tridimensionnelle in vitro pour induire une mutation d'allèle RB1 ou des cellules souches pluripotentes humaines knock-out pour se différencier en un modèle de rétinoblastome humain.
(ZH)
本发明提供一种人视网膜母细胞瘤模型的制备方法,包括:(a)通过对人胚胎干细胞中人视网膜母细胞瘤RB1等位基因进行基因编辑,或将携带RB1等位基因突变的Rb病人的体细胞重编程为人诱导多能干细胞,建立RB1等位基因突变或敲除的人多能干细胞系,进一步筛选及鉴定;(b)利用体外三维视网膜分化体系诱导RB1等位基因突变或敲除的人多能干细胞分化为人视网膜母细胞瘤模型。
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