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Construction method and application of CD226 gene-humanized non-human animal

A CD226, non-human animal technology, applied in chemical instruments and methods, botanical equipment and methods, biochemical equipment and methods, etc., can solve problems such as gene complexity

Active Publication Date: 2020-10-20
BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the differences in physiology and pathology between animals and humans, coupled with the complexity of genes, 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 and application of CD226 gene-humanized non-human animal
  • Construction method and application of CD226 gene-humanized non-human animal
  • Construction method and application of CD226 gene-humanized non-human animal

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0121] Example 1 CD226 Gene Humanized Mice

[0122] Mouse CD226 gene (NCBI Gene ID: 225825, Primary source: MGI: 3039602, UniProt: Q8K4F0, located at positions 89176954 to 89272232 of chromosome 18 NC_000084.6, based on the transcript NM_178687.2 and its encoded protein NP_848802.2 ( SEQ ID NO.1)) and human CD226 gene (NCBI Gene ID: 10666, Primary source: HGNC: 16961, UniProt ID: Q15762, located at positions 69853274 to 69962086 of chromosome 18 NC_000018.10, based on transcript NM_006566.4 and its encoded protein NP_006557.2 (SEQ ID NO.2)) comparison diagram as shown figure 1 shown.

[0123] In order to achieve the purpose of the present invention, the nucleotide sequence encoding human CD226 protein can be introduced into the mouse endogenous CD226 gene locus, so that the mouse expresses human or humanized CD226 protein. Specifically, using gene editing technology, under the control of the regulatory elements of the mouse CD226 gene, the partial sequence of exon 2 to exon ...

Embodiment 2

[0168] Example 2 Preparation of double humanized or multiple double humanized mice

[0169] Double-humanized or multihumanized mouse models can also be prepared by using the method or the prepared CD226 mice. For example, in the aforementioned Example 1, the embryonic stem cells used in the microinjection of blastocysts can be selected from mice containing other genetic modifications such as PD-1, PD-L1, and TIGIT, or they can also be derived from humanized CD226 mice Based on the isolation of mouse ES embryonic stem cells and gene recombination targeting technology, a double-gene or multi-gene modified mouse model of CD226 and other gene modifications was obtained. The homozygous or heterozygous CD226 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 Mendelian law of inheritance, there is a certain probability of obtaining humanized CD226 Heterozygous mice with o...

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PUM

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Abstract

The invention provides a construction method of a CD226 gene humanized modified non-human animal. By utilizing a homologous recombination manner, a nucleotide sequence for encoding a human CD226 protein is introduced into a non-human animal genome. The humanized CD226 protein can be normally expressed in an animal body, and the non-human animal can be used as an animal model for researching humanCD226 signal mechanism and screening drugs for tumor and immune diseases, and has important application value for research and development of new drugs for immune targets. The invention also providesa CD226 chimeric protein, a CD226 chimeric gene, a targeting vector of the CD226 gene, the non-human animal constructed by the construction method, and an application of the non-human animal in the field of biological medicines.

Description

technical field [0001] The invention belongs to the field of animal genetic engineering and genetic modification, and in particular relates to a method for constructing a CD226 genetically modified non-human animal model and its application in the field of biomedicine. Background technique [0002] CD226 is a glycoprotein that belongs to the Ig superfamily and is mainly expressed on the surface of NK cells and T cells, as well as platelets and monocytes. Current studies have shown that the ligands of CD226 are CD112 and CD155, which are expressed in many malignant tumors and virus-infected cells, and their overexpression is related to cancer invasion and metastasis. Therefore, CD226 plays a vital role in anti-tumor immune response and control of viral infection in vivo. Studies have found that changes in the Gly307Ser amino acid of CD226 are also associated with autoimmune diseases, such as type I diabetes, rheumatoid arthritis, and systemic lupus erythematosus. Anti-CD226...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85A01K67/027C07K14/705C12N15/12
CPCA01K67/0278A01K2207/15A01K2217/072A01K2227/105A01K2267/03A01K2267/0393A61K49/0008C07K14/70503C12N15/8509C12N2800/107G01N33/68G01N2333/70503
Inventor 沈月雷郭雅南白阳姚佳维赵磊
Owner BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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