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Construction method and application of CEACAM1 gene humanized non-human animal

A technology for non-human animals and construction methods, applied in chemical instruments and methods, botany equipment and methods, biochemical equipment and methods, etc., capable of solving problems such as genetic complexity

Active Publication Date: 2021-03-16
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, for example, the identity of human and mouse CEACAM1 proteins is only 58%, how can an "effective" humanized animal model be constructed For new drug development is still the biggest challenge

Method used

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  • Construction method and application of CEACAM1 gene humanized non-human animal
  • Construction method and application of CEACAM1 gene humanized non-human animal
  • Construction method and application of CEACAM1 gene humanized non-human animal

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0153]Example 1 CEACAM1 gene sourced mouse

[0154]Mouse CEACAM1 gene (NCBI Gene ID: 26365, Primary Source: MGI: 1347245, UNIPROT: Q925P3, located 2516127 to 251777072 in chromosomes NC_000073.7, based on transcriptional NM_001039185.1 and its encoded protein NP_001034274.1 ( SEQ ID NO: 1)) and human CEACAM1 gene (NCBI Geneid: 634, Primary Source: HGNC: 1814, UNIPROT ID: P13688-1, located 4,2507306 to 42528482 in chromosome NC_000019.10, based on transcript nm_001712. 5 and its encoded protein NP_001703.2 (SEQ ID NO: 2)) Comparison Schematicfigure 1 Indicated.

[0155]To achieve the object of the present invention, the nucleotide sequence encoding the CEACAM1 protein can be introduced in the mouse internal source CEACAM1 locus such that the mouse expresses people or humanized CEACAM1 proteins. Specifically, with gene editing techniques, under the control of the mouse CEACAM1 gene regulatory element, some sequences of mice No. 2 exon partial sequences were replaced by exons molecular seque...

Embodiment 2

[0180]Example 2 Preparation of Dual Source or Multi - Double Source Mouse

[0181]Double source or multi-humanized mouse models can also be prepared using CEACAM1 mice prepared by this method or prepared. As, in the foregoing Example 1, the embryonic stem cells used in the blastocystiovisor can be selected from mice containing other genes such as PD-1, PD-L1, TIGIT, CD226, or in humanization CEACAM1. On the basis of the mice, the mouse model modified by CEACAM1 and other gene-modified bis gene or polygenetic modified mouse model was obtained by isolating mouse ES embryonic stem cells and gene recombinant target techniques. The CEACAM1 mice or heterozygous or heterozygous mice obtained by the method can also be matered with other gene-modified homozygous or heterozygous mice, and the descendants can be screened, according to Mendel's genetic law, can have a certain chance to obtain humanization CEACAM1 Hybridous mice modified with other genetically modified bis gene or polymeric, then m...

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Abstract

The invention provides a construction method of a CEACAM1 gene humanized non-human animal. A nucleotide sequence for encoding human CEACAM1 protein is introduced into a non-human animal genome in a homologous recombination manner, and the humanized CEACAM1 protein can be normally expressed in the body of the animal, can be used as an animal model for human CEACAM1 signal mechanism research and tumor and immune disease drug screening, and has important application value for research and development of new drugs of immune targets. The invention further provides the humanized CEACAM1 protein, a humanized CEACAM1 gene, a CEACAM1 gene targeting vector, the non-human animal obtained with the construction method and application of the non-human animal in the field of biomedicine.

Description

Technical field[0001]The invention belongs to the field of animal genetic engineering and genetic modification, and in particular, the present invention relates to the construction method of CEACAM1 gene humanized non-human animal and its application in the field of biomedical.Background technique[0002]CarcinoemBryonic antigen-related Celladhesion molecule 1, hereinafter referred to as CEACAM1), is also referred to as CD66A or C-CAM, is a transmembrane glycoprotein, belonging to the carcinozyg antigen (CEA) family . CEACAM1 is widely expressed in epithelial cells and vascular endothelial cells, CEACAM1 is also expressed in T cells, B cells, IL-2 activated T cells in granulocytes, mononuclear-macrophages, platelets, B cells, IL-2, and dendritic cells. CEACAM1 has many biological functions that prevent tumors from growing the growth of epithelial cells, inducing apoptosis of epithelial cells, preventing the activation of T lymphocytes, stimulating the proliferation of B lymphocytes, i...

Claims

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Application Information

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IPC IPC(8): C12N15/85A01K67/027C07K14/705C12N15/12C12N5/10A61K49/00
CPCA01K67/0271A01K67/0278A01K2227/105A01K2267/0331A01K2267/0387A61K49/0008C07K14/70503C12N15/8509C12N2800/107
Inventor 赵磊
Owner BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD