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Construction method and application of CD27 humanized mouse model

A humanized and human-derived technology, applied in the construction of CD27 gene-modified humanized animal models, can solve the problem of incomplete testing of antibodies in humans, and achieve the effect of high success rate

Inactive Publication Date: 2020-04-17
GEMPHARMATECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to species differences, the immune checkpoint antibodies screened in mice are not fully tested in humans

Method used

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  • Construction method and application of CD27 humanized mouse model
  • Construction method and application of CD27 humanized mouse model
  • Construction method and application of CD27 humanized mouse model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Establishment of CD27 humanized mouse model

[0043] We used homologous recombination and embryonic stem cell technology to replace the mouse Cd27 gene with human CD27 on mouse ES cells with C57BL / 6 background and BALB / c background, so as to construct a mouse model that can express human CD27. And by mating with PD1 humanized mice with independent property rights, a dual-human mouse model that can express human CD27 and PD1 was obtained.

[0044] 1. Determine the replacement region of the human fragment and the inserted human sequence

[0045] According to the structure and function of human CD27, we selected the extracellular region encoded by the human CD27 gene to replace the extracellular region encoded by the mouse CD27 gene, retained the intracellular region of the mouse, and selected the extracellular region encoded by the human CD27 gene The amino acid sequence (Aa: 1-191) is shown in SEQ ID No: 1.

[0046] MARPHPWWLCVLGTLVGLSATPAPKSCPERHYWAQGKLCCQM...

Embodiment 2

[0078] Example 2: Human CD27 Expression and Immune System Verification in C57BL / 6-hPD1 / hCD27 Mice

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Abstract

The invention provides a method for preparing CD27 gene humanized animals. According to the method, an extracellular region coded by an animal-derived CD27 gene is replaced with an extracellular region coded by a human-derived CD27 gene through homologous recombination; and the intracellular region of the animal-derived CD27 gene is reserved; a humanized CD27 animal model is successfully constructed, and the model can be used for evaluating a CD27 targeted drug or evaluating the curative effect of combination of the CD27 targeted drug and other drugs.

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 humanized animal model of CD27 gene modification based on gene editing technology. Background technique [0002] In recent decades, the development of tumor immunotherapy, especially the discovery and application of immune checkpoint molecules, has subverted the traditional tumor treatment. Immune checkpoint molecules are a class of molecules expressed on the surface of immune cells that can activate or inhibit the function of immune cells. The use of immune checkpoints to treat tumors is to stimulate or inhibit the signal transduction of immune checkpoint proteins through antibodies, and finally activate the immune response of immune cells against tumor cells. Among them, the inhibitory molecules cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and programmed cell death receptor 1 (PD-1) are the two m...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/90C07K19/00A01K67/027A61K49/00
CPCA01K67/0278A01K2207/15A01K2217/072A01K2227/105A01K2267/03A61K49/0008C07K14/70578C07K2319/00C12N15/8509C12N15/907C12N2800/107
Inventor 琚存祥赵静吴丹张明坤王鼎玉侯欢欢高翔
Owner GEMPHARMATECH CO LTD
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