H11 fixed-point knock-in heterozygous mice model of conditional overexpression Spp1 gene and construction method of H11 fixed-point knock-in heterozygous mice model of conditional overexpression Spp1 gene

A technology of mouse model and construction method, which is applied in the field of genetic engineering, can solve the problems of uncontrollable onset time and hinder the understanding of myc gene function, and achieve good controllability

Active Publication Date: 2017-03-08
赣南医学院第一附属医院
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Problems solved by technology

[0006] However, these models also have their limitations: In the case of the Ig heavy chain enhancer-driven SPP1 transgenic mouse (Eμ-myc), overexpression of SPP1 begins at embryonic stage and B cell volume can be observed in postnatal mice Leukemia symptoms such as enlargement hinder the understanding of the function of the myc gene in the process of leukemogenesis caused by SPP1. In this transgenic mouse model, the sustained

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  • H11 fixed-point knock-in heterozygous mice model of conditional overexpression Spp1 gene and construction method of H11 fixed-point knock-in heterozygous mice model of conditional overexpression Spp1 gene
  • H11 fixed-point knock-in heterozygous mice model of conditional overexpression Spp1 gene and construction method of H11 fixed-point knock-in heterozygous mice model of conditional overexpression Spp1 gene
  • H11 fixed-point knock-in heterozygous mice model of conditional overexpression Spp1 gene and construction method of H11 fixed-point knock-in heterozygous mice model of conditional overexpression Spp1 gene

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Embodiment 1

[0032] Example 1 Construction method of H11 site-directed knock-in heterozygous mouse model with conditional overexpression of Spp1 gene

[0033] Include the following steps:

[0034] (1) Obtain gRNA

[0035] gRNA1: 5'-ATGATGGCATCTAATGAGCTTGG-3'

[0036] (2), construction of homologous recombination plasmid

[0037] For a map of homologous recombination plasmids, see figure 2 , which contains all the elements of Table 1. SacI was used to identify the homologous recombination vector, and the identification results were as follows: image 3 As shown, the theoretical band sizes are 5884bp and 14780bp, indicating that the homologous recombination plasmid was successfully constructed.

[0038] Table 1 Elements of Homologous Recombination Plasmids

[0039]

[0040]

[0041] (3), construction of F0 generation mice

[0042] A total of 15 F0 mice were obtained by microinjection of fertilized eggs.

[0043] To identify the genotype of the F0 generation mice, the identific...

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Abstract

The invention discloses an H11 fixed-point knock-in heterozygous mice model of a conditional overexpression Spp1 gene and a construction method of the H11 fixed-point knock-in heterozygous mice model of the conditional overexpression Spp1 gene. The construction method includes obtaining an inducible Spp1 overexpression transgenic vector; obtaining transgenic F0-generation mice; enabling the transgenic F0-generation mice to mate with wild-type mice for passage and line establishment so as to obtain F1-generation mice; selecting the F1-generation mice high in expression in terms of bone marrows, spleens, lymph nodes and thymus glands as a subsequent laboratory strain so as to obtain transgenosis tool mice; enabling the transgenosis tool mice to mate with Cre transgenic mice so as to obtain the H11 fixed-point knock-in heterozygous mice model. The H11 fixed-point knock-in heterozygous mice model, a transgenosis mice model, is an inducible leukemia model characterized in that Spp1 isn't in expression and causes no leukemia in the absence of an inducer and is in expression and causes leukemia in the presence of the inducer, thereby being higher in controllability than a conventional transgenosis mice model and capable of adjusting the quantity of species needing to be maintained according to experiment needs.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and more specifically, the invention relates to a H11 site-specific knock-in heterozygous mouse model with conditional overexpression of SPP1 gene and a construction method thereof. Background technique [0002] Leukemia is a malignant disease that occurs in hematopoietic organs and is characterized by abnormal proliferation and development of white blood cells and their precursor cells in blood and bone marrow. According to the report of the World Health Organization, about 210,000 people died of leukemia in the world in 2000, and 90% of them were adults. The characteristics of the mouse hematopoietic system are similar to those of humans, and mouse leukemia is very similar to human leukemia in many ways. The research and development of mouse leukemia model has become a powerful tool for studying human leukemia pathogenesis, biochemical immune characteristics, pathophysiological cha...

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Application Information

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IPC IPC(8): C12N15/877A01K67/027A01K67/02
CPCA01K67/02A01K67/0278A01K2217/072A01K2227/105A01K2267/0331C12N15/8775
Inventor 张国玺邹晓峰肖观称徐刚夏维
Owner 赣南医学院第一附属医院
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