Preparation method of mouse with specific Raptor knocked out in beta cells during GFP tracing

A specific, β-cell technology, applied in animal husbandry and other fields, can solve problems such as unclear mTORC1β cell identity attributes, glucose intolerance, etc.

Inactive Publication Date: 2019-11-22
RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE +1
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Problems solved by technology

Consistent activation of mTORC1 by overexpression of the brain Ras homolog protein (Rheb) or knockdown of tuberous sclerosis syndrome complex 1 / 2 (TSC1 / TSC2) in vitro resulted in increased β-cell functional mass and insulin secretion, Conversely, mice overexpressing mTOR mutants developed glucose intolerance on a high-fat diet due to a defect in the functionally related factor Pdx1
[0004] It remains unclear whether mTORC1 can regulate β-cell identity properties and whether dysfunction in Raptor knockout mice is independent of glucose metabolism levels

Method used

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  • Preparation method of mouse with specific Raptor knocked out in beta cells during GFP tracing
  • Preparation method of mouse with specific Raptor knocked out in beta cells during GFP tracing
  • Preparation method of mouse with specific Raptor knocked out in beta cells during GFP tracing

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Embodiment 1

[0023] 1. βRap KO GFP mouse build

[0024] SPF (Specific pathogen Free, no specific pathogen) grade Raptor lox / lox C57BL / 6 mice and ROSA26-EGFP f Mixed background mice were purchased from Jackson Laboratory.

[0025]The 8-week-old sexually mature male mice were housed with the female mice, and separated into cages after the 3-week-old mice were born. At the same time, a small amount of nail tissue was cut and numbered for subsequent genotype identification tests. Mate ROSA26-EGFPf with RIP-Cre mice to obtain homozygous mice with RIP-Cre and EGFP sequences. The efficiency of mouse GFP-positive cell sorting was verified by flow sorting technology. Homozygous mice with RIP-Cre and EGFP sequences and Raptor in the same way lox / lox Raptor with homozygous sequences of RIP-Cre and EGFP obtained after mating lox / w mice. The Raptor heterozygous mice were then selfed to obtain Raptor with RIP-Cre and EGFP homozygous sequences lox / lox and Raptor w / w mice, the former is the mouse...

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Abstract

The invention relates to a preparation method of a mouse with specific Raptor knocked out in beta cells during GFP tracing. The method comprises the steps of mating an ROSA26-EGFPf mouse with an RIP-Cre mouse to obtain a homozygous mouse with RIP-Cre and EGFP sequences; then, mating the homozygous mouse with a Raptorlox / lox mouse to obtain a Raptorlox / w mouse with RIP-Cre and EGFP homozygous subsequences; then, selfing the Raptor heterozygous mouse to obtain a Raptorlox / lox mouse with RIP-Cre and EGFP homozygote sequences. The method aims to explain the direct effect of mTORC1 on beta cell identity and maintenance before the onset of diabetes, and provides a further basis for the mTORC1 serving as a drug action target.

Description

technical field [0001] The invention belongs to the field of type 2 diabetes, and in particular relates to a method for preparing mice with specific Raptor knockout in beta cells and GFP tracer. Background technique [0002] Diabetes is the most common metabolic disorder that endangers human health. According to the International Diabetes Federation (IDF), more than 300 million people worldwide suffer from diabetes, and it will increase to 500 million by 2030. The prevalence rate is as high as 11.6%, and only 10% of the patients get better blood sugar control through treatment. Diabetes has become the third largest non-communicable disease after tumors and cardiovascular and cerebrovascular diseases, which has seriously affected national health and economic development. Therefore, it is necessary and urgent to strengthen research on the pathogenesis and therapeutic targets of diabetes . Type 2 diabetes is characterized by pancreatic β-cell dysfunction accompanied by periph...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/02
CPCA01K67/02A01K2217/077A01K2227/105A01K2267/0362
Inventor 宁光汪启迪顾燕云
Owner RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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