Construction method of IL7R gene humanized modified animal model and corresponding application

A technology for constructing methods and animal models, applied in the fields of biomedicine, animal genetic engineering and genetic modification, which can solve problems such as inability to use ordinary mice, inability to identify mice, screening and evaluation of drug efficacy, etc.

Active Publication Date: 2020-10-23
BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because there are significant differences between the human IL7R sequence and the corresponding protein in rodents, for example, the identity of human IL7R and mouse IL7R protein sequences is only 64%, and the antibody that recognizes human IL7R protein usually cannot recognize mouse IL7R, that is, it cannot be used with ordinary Mice to Screen and Evaluate the Efficacy of Drugs Targeting IL7R
In view of the global research and development progress of IL7R-related drugs and the huge application value of targeting this signaling pathway, in order to make pre-clinical trials more effective and minimize the failure rate of research and development, there is an urgent need to develop humanized IL7R-related non-human animal models in this field. However, Due to the differences in physiology and pathology between animals and humans, coupled with the complexity of genes (genetic factors), how to construct an "effective" humanized animal model for new drug development is still the biggest challenge

Method used

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  • Construction method of IL7R gene humanized modified animal model and corresponding application
  • Construction method of IL7R gene humanized modified animal model and corresponding application
  • Construction method of IL7R gene humanized modified animal model and corresponding application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0140] Example 1 IL7R Gene Humanized Mice

[0141] Mouse IL7R gene (NCBI Gene ID: 16197, Primary source: MGI: 96562, UniProtKB: P16872, located at positions 9506159 to 9529941 on chromosome 15 NC_000081.6, based on the transcript NM_008372.4 and its encoded protein NP_032398.3 ( SEQ ID NO.1)) and human IL7R gene (NCBI Gene ID: 3575, Primarysource: HGNC: 6024, UniProtKB: P16871, located at positions 35856891 to 35879603 of chromosome 5 NC_000005.10, based on transcript NM_002185.5 and its The comparison diagram of the encoded protein NP_002176.2 (shown in SEQ ID NO.2) is shown in figure 1 shown.

[0142] In order to achieve the purpose of the present invention, the gene sequence encoding human IL7R protein can be introduced into the mouse endogenous IL7R gene locus, so that the mouse expresses human or humanized IL7R protein. The method of directly inserting the gene sequence containing human IL7R at the endogenous IL7R locus of the mouse can be adopted, such as a DNA sequenc...

Embodiment 2

[0169] Example 2 Double-gene humanized mice and multi-gene humanized mice

[0170] The prepared IL7R mice can also be used to prepare IL7R double humanized or multihumanized mouse models. For example, in the aforementioned Example 1, ES cells used in electroporation were selected from ES cells containing other genetically modified mice, such as PD-1 gene humanized positive clone cells obtained from patent CN201710505554.0, which can further obtain IL7R human Humanized and PD-1 gene-modified double-gene humanized mice. The homozygous or heterozygous IL7R mice obtained by this method can also be mated with other genetically modified homozygous or heterozygous mice, and their offspring can be screened. According to the law of Mendelian inheritance, there is a certain probability of obtaining IL7R humanized and heterozygous mice. Other gene-modified double-gene or polygene-modified heterozygous mice, and then the heterozygotes can be crossed to obtain double-gene or polygene-modi...

Embodiment 3

[0172] Example 3 In vivo drug efficacy verification in animal models

[0173] The IL7R-containing humanized mice prepared by this method can be used to verify the efficacy of the anti-human antibody after establishing a disease model. Taking IL7R single-gene humanized mice and tumor models as examples, mouse tumor cells (such as colon cancer cell MC38) were inoculated subcutaneously in IL7R humanized mice, and were randomly divided into control group or therapy group. Anti-human IL7R antibody was used in the treatment group, and blank solvent was injected in the control group. By measuring the tumor volume and weighing the body weight of the mice, the toxicity, efficacy and combination regimen of the antibody to be tested can be evaluated.

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Abstract

The invention provides a construction method of an IL7R gene humanized modified animal model, a humanized IL7R protein, a humanized IL7R gene, a targeting vector of the IL7R gene and an application ofthe targeting vector in the field of biological medicines. A part of nucleotide sequences for coding the human IL7R protein are introduced into a non-human animal genome in a homologous recombinationmanner, and the animal model can normally express the humanized IL7R protein in vivo, and can be used for human IL7R signal mechanism research, tumor and immune related disease drug screening, and important application value is realized on the research and development of new drugs of immune targets.

Description

technical field [0001] The invention belongs to the field of animal genetic engineering and genetic modification, and in particular relates to a construction method of an IL7R gene humanized transformation animal model and its application in the field of biomedicine. Background technique [0002] Interleukin 7 receptor (IL7R) is a transmembrane receptor belonging to a member of the type I cytokine receptor family. It consists of an α chain (IL7Rα, CD127) and a γ chain (γc, CD132). Specific component; γ chain is a common component of cytokine receptors such as IL2, IL4, IL7, IL9, IL15, IL21, also known as common γ chain, and is an essential component for initiating the expression of IL7R downstream signals. Under physiological conditions, IL7R is an essential factor for the proliferation and differentiation of lymphocytes, and the loss of IL7R on the cell surface can lead to disorders of B-line and T-line lymphocyte production; Acquired mutations can cause acute T lymphoblas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85A01K67/027C07K14/715C12N15/12
CPCA01K67/0278A01K2217/072A01K2227/105A01K2267/03C07K14/7155C12N15/85
Inventor 沈月雷白阳郭朝设张美玲黄蕤周小飞
Owner BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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