Application of RING finger protein rnf20 gene

A gene knockout mouse, fat technology, applied in peptide/protein components, medical preparations containing active ingredients, DNA/RNA fragments, etc., can solve the problem of limited model selection and other issues

Active Publication Date: 2021-07-06
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But options for mouse models of lipodystrophy have long been extremely limited

Method used

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  • Application of RING finger protein rnf20 gene
  • Application of RING finger protein rnf20 gene
  • Application of RING finger protein rnf20 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Preparation of Rnf20 knockout mice

[0070] RNF20 inactivation mutant mice can be obtained by inserting exogenous DNA fragments such as transposons or viruses into the Rnf20 gene sequence, or using chemical mutagenesis methods such as ENU, or using physical mutagenesis methods such as X-rays, or based on embryonic stem cells or CRISPR / Cas9 technology gene targeting and other methods to change the gene sequence, or reduce gene expression.

[0071] In this embodiment, SPF grade Rnf20 is used Flox / Flox Male mice (C57BL / 6J background) and Adiponectin-Cre + (C57BL / 6J background) mice were mated to obtain Rnf20 Flox / + ; Adiponectin-Cre + Rnf20 Flox / Flox Knockout mice (Rnf20 Flox / Flox ; Adiponectin-Cre + , ASKO mice) and littermate control mice (Rnf20 Flox / Flox , WT mice).

[0072] Mice were fed separately under the following conditions: standard commercial mouse feed was purchased from Beijing Keao Xieli Feed Co., Ltd. (4.5% fat, 4% cellulose, 21% protein, ...

Embodiment 2

[0094] Example 2 Knockout of Rnf20 gene leads to reduction of fat deposition

[0095] As the age of Rnf20 knockout mice increased, we detected the body composition of 2-month-old, 6-month-old, and 1-year-old mice by MRI. Calculation of fat and muscle ratios revealed that the fat composition of Rnf20 knockout mice was significantly lower than that of wild-type mice, while the body weight and muscle composition were significantly higher than those of wild-type mice ( figure 2 , A, and image 3 , A). At the same time, we performed physiological anatomy on high-fat and old-age-induced obese mice, and found that Rnf20 knockout mice had severe loss of adipose tissue in different parts ( figure 2 , B and C, and image 3 , B and D), and accompanied by hypertrophy of other tissues such as the liver ( image 3 , D, and Figure 4 , A and B). H&E staining of adipose tissue revealed that the adipocytes of Rnf20 knockout mice were significantly smaller than that of wild-type mice ( ...

Embodiment 3

[0105] Example 3 Knockout of Rnf20 gene leads to hyperinsulinemia

[0106] Insulin is a hormone that balances lipid metabolism and glucose metabolism. Patients with lipodystrophy are often accompanied by symptoms of elevated insulin in serum. In order to detect the level of insulin in Rnf20 knockout mice, we detected the concentration of this hormone in serum of knockout mice by enzyme-linked immunosorbent assay (ELISA). The results showed that serum insulin of 2-month-old, 6-month-old and high-fat-induced 16-week Rnf20 knockout mice was significantly higher than that of wild-type mice ( Figure 4 , C). The experimental results showed that knocking out the Rnf20 gene in mouse adipose tissue would lead to a hyperinsulinemia phenotype. At the same time, we detected the contents of adiponectin and leptin in serum, and found that the contents of these two adipocytokines were significantly lower in ASKO mice than in WT mice at various developmental stages ( Figure 4 , A and B)...

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Abstract

The invention provides the application of the Rnf20 gene of the ring finger protein. The present invention utilizes Cre-loxp system, through Rnf20 Flox / Flox Mouse and Adiponectin‑Cre + The mice were mated, and the adipose tissue-specific Rnf20 gene knockout mouse model was successfully prepared. The present invention uses fat-specific Rnf20 gene knockout mice (Rnf20 Flox / Flox ; Adiponectin‑Cre + ) and control mice (Rnf20 Flox / Flox ) was used as the experimental object, and the function of Rnf20 gene in fat metabolism was studied through animal models induced by age and high-fat diet. The results showed that compared with control mice, fat-specific Rnf20 knockout mice had lipid metabolism disorders, It can simulate the pathogenesis of human lipodystrophy. This model is an ideal model for studying lipodystrophy and has important application value.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of the Rnf20 gene of the ring finger protein. Background technique [0002] White fat plays an important role in maintaining the metabolic balance of the body, and the metabolic disorder caused by its abnormal function has become an important threat to human health. Abnormal fat storage function can mainly lead to obesity and lipodystrophy. Lipodystrophy is characterized by complete or partial loss of adipose tissue. Congenital generalized lipodystrophy is a rare autosomal recessive disease, clinically manifested as almost complete loss of adipose tissue throughout the body, accompanied by metabolic abnormalities, such as insulin resistance, impaired glucose tolerance, fatty liver, dyslipidemia, Serum leptin, adiponectin levels decreased, etc. Therefore, adipose tissue is considered to be one of the most important endocrine organs in animals. [0003] Histone post-...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/00C12N15/52A61K38/17A61K45/00A61P3/04A61P3/00
CPCA61K38/17A61K45/00A61P3/00A61P3/04C12N9/93C12Y603/02019
Inventor 王彦芳梁小娟陶聪王超王亚君
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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