Method for constructing kidney persistent innate macrophage infiltration rat model

A macrophage and mouse model technology, applied in biochemical equipment and methods, animal husbandry, and the use of vectors to introduce foreign genetic material, etc., can solve the problem of single factor analysis of kidney function and cannot be used as a research model of kidney intrinsic macrophages To achieve the effect of avoiding non-selective gene overexpression and non-selective cell infiltration, improving specificity and reducing influence

Active Publication Date: 2018-08-31
武汉麦考津生物科技有限公司
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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 intuit

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  • Method for constructing kidney persistent innate macrophage infiltration rat model
  • Method for constructing kidney persistent innate macrophage infiltration rat model

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Embodiment Construction

[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 shown in 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 direction...

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Abstract

The invention relates to a method for constructing a kidney persistent innate macrophage infiltration rat model. The method includes the step of specifically expressing an IL34 gene in a rat kidney tubules cell. Preferentially, a totipotential cell contains a Cre recombinase expression frame and an IL34 condition expression frame driven by a kidney tubules tissue specific promoter at the same time, and the IL34 condition expression frame comprises a target gene expression promoter, a transcription barrier segment with loxp sequences in the same direction at the two ends, and an IL34 gene coding frame. The method has the advantages that by constructing a kidney tubules height expression IL34 rat, the kidney rat model with innate macrophages as the main infiltration characteristic is successfully constructed, unselective gene over-expression and unselective cell infiltration can be effectively avoided, the influences on the overall state of important organs outside a kidney and an organism are reduced, and the specificity of the kidney innate macrophage infiltration can be effectively improved.

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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IPC IPC(8): C12N15/85A01K67/027
CPCA01K67/0278A01K2227/105A01K2267/0306C12N15/8509
Inventor 曾锐廖文慧
Owner 武汉麦考津生物科技有限公司
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