Method for constructing mouse model capable of specifically knocking out IKKalpha gene in hippocampus region, targeting vector and kit

Abstract

The invention discloses a method for constructing a mouse model capable of specifically knocking out an IKKalpha gene in a hippocampus region, a targeting vector and a kit. The IKKalpha gene is knocked out from specific mouse tissues through utilizing a Cre-LoxP recombination system, so that a transgenic mouse with the knocked-out IKKalpha gene is established, and an NF-omicronB signal channel and an acting mechanism on diseases, particularly tumors, are researched on the whole level of animals. Meanwhile, in order to determine and monitor whether specific knockout of the IKKalpha gene in the hippocampus region causes influences on the behavioral science of the mouse, red luciferase RFP720 is connected into a transgenic carrier. A gene targeting technology is adopted, and expression of the endogenous IKKalpha gene in the hippocampus region of the mouse is reduced based on iRFP720 near-infrared fluorescent protein and a Cre-LoxP gene knockout system, so that living body imaging of the transgenic mouse is monitored through an animal imaging system.

Description

technical field [0001] The invention belongs to the technical fields of imaging, pathology, physiology, genetics, molecular biology and behavior. It specifically relates to a method for constructing a mouse model for specifically knocking out the IKKα gene in the hippocampus region, as well as a targeting vector and a kit. Background technique [0002] IKKα is a regulatory subunit that constitutes the IKK complex, which can affect the activation of the NF-κB signaling pathway. Studies on glioma (a tumor that occurs in the brain) have shown that NF-κB and other signaling pathways related to inflammation are abnormally activated, but the mechanism of IKKα in brain tumors is currently unclear. Whether it will affect the occurrence and development of brain tumors is not yet clear. At the same time, as an inflammation-related molecule, whether it will affect its behavioral function after complete knockout in the normal mammalian hippocampus is not clear. Therefore, using animal...

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Problems solved by technology

[0006] 2. If the two LoxP sites are located on one DNA strand, but in opposite directions, Cre recombinase can cause sequence inversion between the two LoxP sites;

iRFP720 is a near-infrared fluorescent protein gene, which is isolated from bacterial photoreceptor cells and can be used for genetic engineering editing to construct tags for targeting vectors. At present, there is a lack of research on iRFP720 as a near-infrared red fluorescent tag to construct animal models

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