Construction method of CNN3 gene knockout mouse model based on Cre-FloxP system

A technology for gene knockout mice and construction methods, applied in the field of genes, can solve the problems of low efficiency of mouse models

Pending Publication Date: 2021-11-02
FIRST AFFILIATED HOSPITAL OF KUNMING MEDICAL UNIV
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[0006] The embodiment of the present invention provides a method for constructing a CNN3 gene knockout mouse model based on the Cre-FloxP sy

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  • Construction method of CNN3 gene knockout mouse model based on Cre-FloxP system
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  • Construction method of CNN3 gene knockout mouse model based on Cre-FloxP system

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Embodiment Construction

[0021] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are some of the embodiments of the present invention, but not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0022] Each raw material and equipment are introduced in the embodiment:

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Abstract

The invention discloses a construction method of a CNN3 gene knockout mouse model based on a Cre-FloxP system, which comprises the following steps: respectively inserting FloxP gene segments at two sides of a second exon of a CNN3 gene, constructing a Cas9/gRNA target and a homologous recombination template, carrying out microinjection on the homologous recombination template and donor DNA into a fertilized egg of a mouse to obtain a Cnn3-FloxP mouse, and carrying out Cre-FloxP system-based CNN3 gene knockout mouse model; after mating with a female mouse, conducting selfing to obtain a stably inherited homozygote CNN3fl/fl mouse; after the homozygote CNN3fl/fl mouse and a Cre mouse with tissue specific expression are copulated, conducting selfing, and screening out the mouse with the genotype of Cre +/-/CNN3fl/fl through gene recognition. The invention relates to the technical field of gene design, can efficiently construct a mouse with specific tissue or specific cell CNN3 gene knockout, and greatly improves the construction efficiency and the construction success rate of the mouse model with the selective CNN3 knockout function.

Description

technical field [0001] The invention relates to the field of gene technology, in particular to a method for constructing a CNN3 gene knockout mouse model based on the Cre-FloxP system. Background technique [0002] Calponin protein is calcium / calmodulin and actin binding protein, there are 3 subtypes: calponin-1 (basic calponin), calponin-2 (neutralcalponin) and calponin-3 (acid calponin). Calponin-3 is the only subtype distributed in the central nervous system in the calponin family, and it is mainly concentrated in astrocytes and neurons in the brain. [0003] The regulatory gene CNN3 of calponin-3 protein is located on chromosome 1p21-22. CNN3 was originally found to regulate neural tube morphogenesis in embryonic development. Systemic knockout of CNN3 in mice results in embryonic and neonatal death due to defects in central nervous system development. Subsequent studies provided evidence for the function of CNN3 beyond the domain of neural growth and development. Cal...

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Application Information

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IPC IPC(8): C12N15/113C12N9/22C12N15/85C12N15/90A01K67/027
CPCC12N15/113C12N9/22C12N15/8509C12N15/907A01K67/0276A01K2207/15A01K2217/075A01K2267/03A01K2227/105
Inventor 韩雁冰余彦萤周亮杜孙兵周启心杨飞陈丽凌徐亭晚陆地边立功郭家智
Owner FIRST AFFILIATED HOSPITAL OF KUNMING MEDICAL UNIV
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