Construction method of CD155 gene humanized non-human animal and application

A non-human animal and construction method technology, which is applied in the field of CD155 genetically modified non-human animal model construction, can solve the problem that animal models cannot reflect the real situation of the human body well

Active Publication Date: 2021-03-09
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 the physiological structure and metabolic system between animals and humans, traditional animal models cannot well reflect the real conditions of the human body. It is an urgent need for the biomedical industry to establish disease models in animals that are closer to the physiological characteristics of humans

Method used

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  • Construction method of CD155 gene humanized non-human animal and application
  • Construction method of CD155 gene humanized non-human animal and application
  • Construction method of CD155 gene humanized non-human animal and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0129] Example 1 CD155 Gene Humanized Mice

[0130]Mouse CD155 gene (NCBI Gene ID: 52118, Primary source: MGI: 107741, UniProt: Q8K094, located at positions 19637503 to 19655068 of chromosome 7 NC_000073.7, based on the transcript NM_027514.2 and its encoded protein NP_081790.1 ( SEQ ID NO: 1)) and human CD155 gene (NCBI Gene ID: 5817, Primarysource: HGNC: 9705, UniProt ID: A0A0C4DG49, located at positions 44643910 to 44666162 of chromosome 19 NC_000019.10, based on transcripts NM_006505.5 and The comparison diagram of its encoded protein NP_006496.4 (SEQ ID NO: 2)) is shown in figure 1 shown.

[0131] In order to achieve the purpose of the present invention, the nucleotide sequence encoding human CD155 protein can be introduced into the mouse endogenous CD155 gene locus, so that the mouse expresses human or humanized CD155 protein. Specifically, using gene editing technology, under the control of mouse CD155 gene regulatory elements, the partial sequence of exon 2 to exon 6...

Embodiment 2

[0162] Example 2 Preparation of double humanized or multiple double humanized mice

[0163] Double-humanized or multi-humanized mouse models can also be prepared by using the method or the prepared CD155 mice. For example, in the aforementioned Example 1, the embryonic stem cells used for blastocyst microinjection can be selected from mice containing other genetic modifications such as PD-1, PD-L1, TIGIT, and CD226, or they can also be derived from humanized CD155 On the basis of mice, use the isolation of mouse ES embryonic stem cells and gene recombination targeting technology to obtain CD155 and other genetically modified double-gene or multi-gene modified mouse models. The homozygous or heterozygous CD155 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 CD155 Heterozygous mi...

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Abstract

The invention provides a construction method of a CD155 gene humanized non-human animal. A nucleotide sequence for encoding human CD155 protein is introduced into a non-human animal genome in a homologous recombination manner; the humanized CD155 protein can be normally expressed in the animal; and the animal can be used as an animal model for human CD155 signal mechanism research and tumour and immune disease drug screening. The important application value is realized on the research and development of new drugs of immune targets. The invention also provides humanized CD155 protein, a humanized CD155 gene, a targeting vector of the CD155 gene, a non-human animal obtained by the construction method, and 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 CD155 genetically modified non-human animal model and its application in the field of biomedicine. Background technique [0002] CD155, also known as PVS, HVED, NECL5, TAGE4 or Necl-5, is a Nectin-like type I transmembrane glycoprotein, which belongs to the PVR-related (PRR) family under the immunoglobulin superfamily, and is an adhesion that promotes cell binding Protein, which can bind to poliovirus (PV for short) and allow it to enter susceptible cells, so it is more commonly called PVR (Poliovirus Receptor, poliovirus receptor). CD155 is expressed in monocytes, macrophages, endothelial cells, epithelial cells, CD34+ thymocytes and neurons, but it is highly expressed in most tumor cells, including lung cancer, breast cancer, ovarian cancer and melanoma. Studies have shown that CD155 plays an important r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C07K14/705C12N15/12A01K67/027A61K49/00
CPCA01K67/0271A01K67/0278A01K2227/105A01K2267/0331A61K49/0008C07K14/70503C12N15/8509
Inventor 张淑金
Owner BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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