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Construction method and application of conditional ivermectin receptor ivmr transgenic mouse model

A technology of transgenic mice and ivermectin, which is applied to other methods of inserting foreign genetic materials, preparations for in vivo tests, pharmaceutical formulas, etc., to achieve the effect of stable half-life and convenient comparison between before and after experiments.

Active Publication Date: 2017-02-22
TONGJI UNIV
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Problems solved by technology

[0003] The purpose of the present invention is to provide a conditional type (or known as the conditional type) that can be better used for the research of the influence of neuron activity on animal behavior in order to address the deficiencies of the existing IVMR transgenic mice lacking ivermectin receptors. Inducible) ivermectin receptor IVMR transgenic mouse model and its construction method and application

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  • Construction method and application of conditional ivermectin receptor ivmr transgenic mouse model

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Embodiment

[0025] 1. Construction of targeting vectors for IVMR transgenic mice

[0026] The plasmid containing the IVMR-2A-GFP gene sequence was donated by Professor Joseph W. Lynch of the University of Queensland, Australia, and the Rosa26-CAG targeting vector was purchased from the Addgene website in the United States. Using the general molecular cloning method, insert the IVMR-2A-GFP gene sequence (GFP is green fluorescent protein, 2A is a self-digesting sequence, which is convenient for separating IVMR and GFP proteins) into the back of the STOP site of the Rosa26 targeting vector, replacing the original The tdTomato sequence in the vector, so as to construct the targeting vector successfully containing the IVMR gene, such as figure 1 As shown in B, the 5' homology arm (5'arm) and the 3' homology arm (3'arm) used in the targeting vector (B) are located at the first and second extrinsics of the wild-type Rosa26 gene locus (A) between exons. Homologous recombination inserts the gene...

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Abstract

The invention relates to a construction method and an application of a conditional ivermectin receptor (IVMR) transgenic mouse model, and belongs to the technical fields of conditional animal model construction method and application. Under the action of a Cre recombinase, all cells (especially cerebral neurons) of the mouse model can conditionally express the ivermectin receptor IVMR, and the excitability of the neurons expressing the ivermectin receptor IVMR can be down-regulated by applying a drug ivermectin (IVM). Compared with the prior art, the technical method (including a drug administration mode, and targetedly using a transgenic mouse for specific expression of Cre) does not has any brain damage on the mouse. Influence of the used drug ivermectin IVM on animals has a sufficiently long half-life period, and animal behavioral experiments are suitably carried out. In addition, the half-life period is relatively stable, the experiments can be again carried out after drug failure, and before-and-after comparison experiments are conveniently carried out on the same animal.

Description

technical field [0001] The invention relates to a construction method and application of a conditional transgenic animal model, in particular to a construction method of a conditional ivermectin receptor IVMR transgenic mouse model and its application in neuron function research. Background technique [0002] A key question in the field of neuroscience research is the causal relationship between the activity (excitability) of specific neurons and behavior. For this problem, many new methods have been developed internationally, such as molecular manipulation methods of synaptic transmission, optogenetics methods and ligand-gated ion channel methods. In particular, ligand-gated ion channels have been developed for inducible and reversible modulation of neuronal activity. For example, Drosophila allatostatin receptors, when activated by allatostatin, can inhibit the generation of action potentials in neurons, a method that has been used in mammalian brains. However, the molec...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/89A01K67/027A61K49/00
Inventor 丁玉强胡玲张玲黄缨张磊宋宁宁
Owner TONGJI UNIV
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