Method for constructing heterozygote mouse model of Amh gene fixed-point knock-in 2A-Cre and application thereof

A technology of mouse model and construction method, applied in other methods of inserting foreign genetic materials, stably introducing foreign DNA into chromosomes, DNA/RNA fragments, etc., can solve problems that restrict the progress of ovarian development and function research

Active Publication Date: 2019-06-28
SHANGHAI GERIATRIC INST OF CHINESE MEDICINE
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

At present, there is no specific Cre transgenic tool mouse in ovarian granulosa c

Method used

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  • Method for constructing heterozygote mouse model of Amh gene fixed-point knock-in 2A-Cre and application thereof
  • Method for constructing heterozygote mouse model of Amh gene fixed-point knock-in 2A-Cre and application thereof
  • Method for constructing heterozygote mouse model of Amh gene fixed-point knock-in 2A-Cre and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0120] 1. Experimental animals

[0121] 5-week-old healthy C57BL / 6 mice (weight 18±2g), 100 male and 100 each, provided by Shanghai Southern Model Biotechnology Co., Ltd., can reach the 6-week-old required by the experiment after one week of isolation; 60 healthy ICR mice aged 6 to 8 weeks, half male and half female, weighing 24±2g, were purchased from Shanghai Southern Model Biotechnology Co., Ltd., and the mice were fed in an SPF-grade environment, strictly in accordance with the management regulations of the animal room , to ensure that the daily temperature is 22°C, the humidity is 40% to 60%, and the light time and dark time in the animal room are half and half. All related operations of animal experiments strictly follow the relevant management regulations of the animal center.

[0122] 2. Experimental reagents

[0123]Cas9Nuclease (NEB USA); Premix TaqTM PCR Enzyme (TAKARA Japan); DNA Purification and Recovery Kit (Tianjin TIANGEN China); Cas9mRNA for Injection (Beijin...

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Abstract

The invention discloses a method for constructing a heterozygote mouse model of an Amh gene fixed-point knock-in 2A-Cre and an application thereof. According to the method, the heterozygote mouse model can be obtained, and an effective experimental animal model is provided for the research of POF pathogenesis, the research and development of drugs and the evaluation of drug effect. The invention provides the application of the heterozygote mouse model in screening and preparing drugs for detecting/treating reproductive development diseases, drugs for detecting/treating ovarian diseases and drugs for aging and apoptosis diseases of ovarian granulosa cells. The invention provides the method for constructing the heterozygote mouse model for the first time, and no domestic and foreign literature reports are found before. The model has good medical clinical application prospect, has great application value in preparing medicaments for detecting/treating ovarian diseases, and has great social benefit.

Description

technical field [0001] The invention relates to biotechnology, in particular to a method for constructing a heterozygous mouse model in which Amh gene is knocked into 2A-Cre and its application. Background technique [0002] Ovarian granulosa cells are important for follicle development and maturation as supporting cells. However, the mechanism of action of ovarian granulosa cells is still unclear. AMH, the anti-Müllerian hormone, is secreted by ovarian granulosa cells and has the function of regulating the development and differentiation of follicle cells. At present, there is no specific Cre transgenic tool mouse in ovarian granulosa cells, which restricts the progress of research on ovarian development and function. Amh gene: Anti-Mullerian hormone (AMH) is one of the members of the transforming growth factor β superfamily. It is a dimeric glycoprotein composed of two identical 70KD subunits linked by disulfide bonds. The relative molecular mass is 140KD; the human AMH...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/90A01K67/027A61K49/00
Inventor 刘特郑锦陈川郁志华林佳佳
Owner SHANGHAI GERIATRIC INST OF CHINESE MEDICINE
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