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Construction method and application of amh gene site-specific knock-in 2a-cre heterozygous mouse model

A mouse model, construction method technology, applied to other methods of inserting foreign genetic material, stably introducing foreign DNA into chromosomes, DNA/RNA fragments, etc., can solve the problems that restrict the progress of ovarian development and function research, etc. To achieve the effect of good medical clinical application prospect, great social benefit and great application value

Active Publication Date: 2022-03-15
SHANGHAI GERIATRIC INST OF CHINESE MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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

Method used

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  • Construction method and application of amh gene site-specific knock-in 2a-cre heterozygous mouse model
  • Construction method and application of amh gene site-specific knock-in 2a-cre heterozygous mouse model
  • Construction method and application of amh gene site-specific knock-in 2a-cre heterozygous mouse model

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Experimental program
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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 heterozygous mouse model in which the Amh gene is knocked in 2A-Cre and its application. The heterozygous mouse model can be obtained through the construction method of the present invention, which provides an effective experimental animal model for research on the pathogenesis of POF, drug development and drug efficacy evaluation. The present invention provides the application of Amh gene site-directed knock-in 2A-Cre heterozygous mouse model in the screening and preparation of drugs for detecting / treating reproductive and developmental diseases, drugs for detecting / treating ovarian diseases, and drugs for aging and apoptosis of ovarian granulosa cells . The present invention provides for the first time a method for constructing a heterozygous mouse model in which the Amh gene is knocked into 2A-Cre, which has not been reported in any domestic or foreign literature before. The invention has a good prospect of medical clinical application, has great application value in the preparation of drugs for detecting / treating ovarian diseases, and has great social benefits.

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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Patent Type & Authority Patents(China)
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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