A method for constructing a mouse model of persistent intrinsic macrophage infiltration in the kidney

A macrophage and mouse model technology, applied in biochemical equipment and methods, animal husbandry, recombinant DNA technology, etc., can solve the problems of inability to single-factor analysis of kidney function, inability to be used as a research model of renal intrinsic macrophages, etc.

Active Publication Date: 2021-08-20
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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Problems solved by technology

[0004] 1) Since this mouse is not a kidney-specific IL34 knockout mouse, let alone a kidney-specific IL34 overexpression mouse, it involves the comprehensive effects of multiple organs and systems, and there is an interaction between each system, so it is impossible to intuitively understand the kidney. One-way analysis of the effect;
[0005] 2) This mouse is also involved in the comprehensive effects of other immune organs, immune cells and other macrophage subtypes, so it cannot be used as a research model for intrinsic macrophages in the kidney

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  • A method for constructing a mouse model of persistent intrinsic macrophage infiltration in the kidney
  • A method for constructing a mouse model of persistent intrinsic macrophage infiltration in the kidney

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[0023] The principles and features of the present invention are described below in conjunction with the accompanying drawings, and the examples given are only used to explain the present invention, and are not intended to limit the scope of the present invention.

[0024] 1. Construction of targeting vector

[0025] The schematic diagram of the targeting carrier is as follows: figure 1 As shown, from 5' to 3' are CAG promoter, loxP sequence, STOP sequence, loxP sequence, IL34 coding frame (Gene id 76527, GenBank), IRES, EGFP, WPRE and PA.

[0026] Among them, the CAG promoter is a constitutive strong promoter, driving the transcription of downstream sequences;

[0027] The Loxp sequence is composed of two 13bp inverted repeat sequences and an 8bp sequence in the middle. The 8bp spacer sequence also determines the direction of LoxP. In this case, the two Loxp sequences have the same direction. When the two Loxp sites are located on the same chromosome and have the same direct...

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Abstract

The present invention relates to the present invention provides a method for constructing a mouse model of persistent innate macrophage infiltration in the kidney, which includes the step of expressing IL34 gene specifically in mouse renal tubular cells. Preferably, the totipotent cells have both a Cre recombinase expression cassette driven by a renal tubular tissue-specific promoter and an IL34 conditional expression cassette, the IL34 conditional expression cassette includes a target gene expression promoter, and two ends with Transcription hindered fragment of loxp sequence and coding frame of IL34 gene. The beneficial effect of the present invention is: by constructing the mice with highly expressed IL34 in renal tubules, a kidney mouse model characterized by infiltration of intrinsic macrophages is successfully constructed, which can effectively avoid non-selective gene overexpression and non-selective cell Infiltration can reduce the impact on vital organs outside the kidney and the overall state of the body, and can also effectively improve the specificity of infiltration of intrinsic macrophages in the kidney.

Description

technical field [0001] The invention relates to the field of mouse disease models, in particular to a method for constructing a mouse model of persistent intrinsic macrophage infiltration in the kidney. Background technique [0002] Macrophages are highly heterogeneous. During the resting period of the kidney, they maintain tissue homeostasis and tissue repair; in pathological conditions, they promote renal fibrosis. Most previous studies simply divided macrophages into M1 (inflammatory macrophages) and M2 (reparative macrophages), ignoring the phenotype, function, connotation and environmental response of macrophages. This complexity, while ignoring the phenomenon of M1 / M2 functional crossover, has led to ambiguity about the role of macrophages in kidney disease. Therefore, more and more researchers now advocate that macrophages in tissues can be divided into peripheral macrophages and intrinsic macrophages (Tissue-resident macrophages) according to their origin. Peripher...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/85A01K67/027
CPCA01K67/0278A01K2227/105A01K2267/0306C12N15/8509
Inventor 曾锐廖文慧
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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