Construction method of FNDC5 gene knock-out rat model, and application

A technology of gene knockout and rat model, applied in the construction of FNDC5 gene knockout rat model, field of gene knockout

Inactive Publication Date: 2020-07-10
CHINA INST OF SPORT SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is no FNDC5 gene knockout rat model at home and abroad. By establishing this model animal, it is convenient to study the function of this gene and Irisin in animals.

Method used

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  • Construction method of FNDC5 gene knock-out rat model, and application
  • Construction method of FNDC5 gene knock-out rat model, and application
  • Construction method of FNDC5 gene knock-out rat model, and application

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Embodiment 1

[0029] 1. Design of gene knockout model rats

[0030] like figure 1 As shown, the FNDC5 gene is on the positive strand of chromosome 5, with a total length of about 7kb. Gene ID: 260327. like figure 2 As shown, the gene has 1 transcript, and this design refers to transcript 201. The FNDC5 gene sequence is shown in SEQ ID NO.8.

[0031] Analysis of the FNDC5 gene structure, selection to delete Exon2-6. Two sgRNAs were designed on Intron1 and 3'UTR respectively, resulting in about 4kb base deletion, so as to achieve the purpose of FNDC5 knockout. Model rats were prepared using the CRISPR / Cas9 system. Knockout strategies such as image 3 shown.

[0032] 2. Preparation of gene knockout rat model

[0033] 2.1 Cas9 / sgRNA design and activity detection

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Abstract

The invention discloses a construction method of an FNDC5 gene knock-out rat model, and an application. The invention discloses two sgRNAs in accordance with a FNDC5 gene of a rat, and a correspondingtarget gene sequence, and the two sgRNAs are respectively designed on FNDC5 genes Intron1 and 3'UTR. The sgRNAs can be applied to construction of a rat animal model having insulin resistance and glycometabolism disorders. The invention provides a simple reliable economic animal model making method having insulin resistance and glycometabolism disorders.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to gene knockout technology, in particular to the construction of FNDC5 gene knockout rat model. Background technique [0002] Insulin resistance (IR) mainly manifests as glucose metabolism disorder and lipid metabolism disorder, leading to glucotoxicity and lipotoxicity, which in turn aggravates lipid metabolism disorder, forming a vicious circle. Irisin plays an important role in the regulation of glucose and lipid metabolism in the body. Exercise induces the expression of muscle fibronectin type III domain (FNDC) 5 by up-regulating the body's oxisome proliferator-activated receptor gamma coactivator 1-α (PGC-1α), and FNDC5 is hydrolyzed by proteolytic enzymes A small peptide, irisin, is produced. Irisin has potential multiple positive effects on glucose homeostasis and insulin sensitivity in the body, mainly in promoting energy expenditure, glucose uptake and glycogenolysis, and reduc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/90A01K67/027
CPCA01K67/0276A01K2217/075A01K2227/105A01K2267/0362C07K14/78C12N15/113C12N15/907C12N2310/20
Inventor 赵杰修房国梁覃飞崔书强徐旻霄瞿超艺董亚南
Owner CHINA INST OF SPORT SCI
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