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Construction method and application of Dkk3 gene mouse model

A mouse model, construction method technology, applied in genetic engineering, biochemical equipment and methods, recombinant DNA technology, etc.

Active Publication Date: 2021-05-25
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no clinically effective inner ear-targeted drug available for the treatment of sensorineural hearing loss.

Method used

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  • Construction method and application of Dkk3 gene mouse model
  • Construction method and application of Dkk3 gene mouse model
  • Construction method and application of Dkk3 gene mouse model

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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 only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.

[0022] like Figure 1-4 As shown, in an example of the present invention, a method for constructing a Dkk3 gene mouse model is disclosed, comprising the following steps: Step 1: cultivating OC-1 cells; knocking down the Dkk3 gene in the OC-1 cells with siRNA , to detect the knockdown efficiency; step 2: take the cochlea of ​​newborn Lgr5-EGFP mice, and digest them into single cells; sort out Lgr5+ cells from the Lgr5-EGFP cell suspension; step 3: knockdown efficien...

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Abstract

The invention discloses a construction method and application of a Dkk3 gene mouse model, and belongs to the technical field of biology. The invention relates to a construction method of a Dkk3 gene mouse model. The construction method comprises the following steps: step 1, culturing OC-1 cells; knocking down the Dkk3 gene in the OC-1 cell by siRNA, and detecting the knock-down efficiency; 2, taking the cochlea of a newborn Lgr5-EGFP mouse, and digesting the cochlea into single cells; separating the Lgr5+ cells from the Lgr5-EGFP cell suspension liquid; step 3, knocking down the Dkk3 gene in the Lgr5 + cell by siRNA with the optimal knocking-down efficiency in the OC-1 cell; and 4, carrying out a cell balling test, and observing and quantifying the number and diameter of spheres to evaluate the proliferation capacity of the Lgr5 + cells.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing a Dkk3 gene mouse model and its application. Background technique [0002] With the development and aggravation of adverse factors such as population aging, noise, virus infection, ototoxic drug abuse, and environmental pollution, the number of deaf and hearing-impaired people is gradually increasing. Deafness has become a global health problem affecting society, politics and economy, and sensorineural deafness accounts for about 63% of deaf patients. However, there is currently no clinically effective inner ear-targeted drug available for the treatment of sensorineural hearing loss. Since mammalian cochlear hair cells cannot regenerate, how to repair and regenerate hair cells after injury, so as to fundamentally treat sensorineural deafness, has become the focus of research in the field of hearing in recent years. It is also of great significance to provi...

Claims

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Application Information

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IPC IPC(8): C12N15/113A01K67/027
CPCC12N15/113A01K67/0276C12N2310/14A01K2207/15A01K2217/075A01K2227/105A01K2267/035Y02A50/30
Inventor 柴人杰李益歌张莎莎程诚高下
Owner SOUTHEAST UNIV
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