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Gene editing method for humanized immune system mouse and application of gene editing method

A technology for immune system and immunodeficiency mice, applied in chemical instruments and methods, biochemical equipment and methods, and other methods of inserting foreign genetic materials, to achieve the goals of improving success rate and reconstruction ratio, prolonging lifespan, and good positive rate Effect

Active Publication Date: 2021-02-02
澎立生物医药技术(上海)股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This technology can solve the common problem of excessively high levels of human cytokines and myeloid immune cells in mice with human-derived immune cells, and can effectively prolong the lifespan of mice while ensuring the human life

Method used

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  • Gene editing method for humanized immune system mouse and application of gene editing method
  • Gene editing method for humanized immune system mouse and application of gene editing method
  • Gene editing method for humanized immune system mouse and application of gene editing method

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Embodiment Construction

[0070] The technical solutions of the present invention are clearly and completely described and illustrated below in conjunction with the embodiments.

[0071] Based on the severe immunodeficiency mouse NPLG mouse (CN202010373356.5) previously developed by the inventor, the CRISPR Cas9 technology was further used on the basis of the mouse to treat IL3, GM-CSF (CSF2) and Kitlg of the NPLG mouse In situ knock-in replacement of human genes.

[0072] 1. Gene knock-in site determination and donor vector construction

[0073] 1. Distribution of each gene

[0074] The mouse IL3 gene (Ensembl database transcript: MGP_NODShiLtJ_T0028883.1) is located on mouse chromosome 11. Five exons have been identified, of which the first exon is the ATG start codon, and the fifth exon is the TAA stop codon.

[0075] The human IL3 gene (NCBI database reference sequence: NM_000588.4) is located on human chromosome 5. Five exons have been identified, of which exon 1 is the ATG start codon and exo...

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Abstract

The invention relates to a preparation method of a humanized NPLG IL3 GM-CSF Kitlg mouse. The method comprises the step of performing in-situ knock-in replacement on humanized genes of IL3, GM-CSF (CSF2) and Kitlg genes of an NPLG mouse with severe immunodeficiency. On the basis of the mouse obtained by the method, the humanized immune system mouse is further prepared, and the humanized immune system mouse has higher humanized cell reconstruction proportion and success rate and longer service life, and can better exert the application value.

Description

technical field [0001] The present invention relates to the technical field of animal genetic engineering and genetic modification, in particular to a gene editing method for humanized immune system mice and its application. Background technique [0002] In biomedical experiments, in order to better study and treat human diseases, it is often necessary to use a variety of different species of animals, such as rats and mice, to simulate human diseases and conduct related research in them. However, there are often large species differences between rodents and humans, and the experimental results obtained from animal models based on rats and mice often have a large gap with the results in the population, and cannot truthfully reflect the effects of drugs in humans. However, primate applications are limited by cost, animal welfare and ethics. In this case, in order to better study the function of human blood and immune system, mice with humanized immune system came into being. ...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/90C12N15/62C12N15/12A01K67/027A61K49/00
CPCA01K67/0278A01K2207/15A01K2217/072A01K2227/105A01K2267/0387A61K49/0008C07K14/475C07K14/535C07K14/5403C07K2319/00C12N15/8509C12N15/907
Inventor 杨楠仝莉黄哲夫陈洁罗艳段继峰
Owner 澎立生物医药技术(上海)股份有限公司