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Method for constructing and identifying systemic Plin1 gene knockout animal model

A technology of animal models and construction methods, applied in the biological field, to achieve the effects of convenient pathogenesis, improved knockout efficiency, and economical animal models

Active Publication Date: 2020-11-27
SHANXI MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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However, there are no reports on the construction and identification of systemic Plin1 gene knockout mouse animal models

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  • Method for constructing and identifying systemic Plin1 gene knockout animal model
  • Method for constructing and identifying systemic Plin1 gene knockout animal model
  • Method for constructing and identifying systemic Plin1 gene knockout animal model

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Embodiment Construction

[0038] The specific implementation manners of the present invention will be further described below. In order to facilitate those skilled in the art to understand the present invention, it should be clear that the present invention is not limited to the scope of specific embodiments. For those skilled in the art, as long as various changes are within the spirit and scope of the present invention defined and determined by the claims, These changes are obvious, and all inventions and creations using the concept of the present invention are included in the protection.

[0039] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The nomenclature and experimental procedures used herein are those of well known and commonly used in the art. All manipulations using standard techniques are generally performed in accordance with the instrument consumabl...

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Abstract

The invention belongs to the technical field of biology, and provides a method for constructing and identifying systemic Plin1 gene knockout animal model. The construction method comprises the following steps: designing sgRNA sequences aiming at a front sequence and a rear sequence of an exon 2 of a Plin1 gene, constructing a Plin1 gene knockout vector, and carrying out in-vitro transcription to obtain sgRNA; and mixing the sgRNA with Cas9 protein, and then performing microinjection into fertilized eggs of a mouse to obtain a systemic Plin1 gene knockout mouse. The method has the advantages that: a pair of the sgRNA sequences with the highest score is screened out, and the success rate of the project is guaranteed; the large-fragment knockout of the exon 2 of the Plin1 gene is realized, and the knockout efficiency is improved; and three primers are designed for the knockout area, various genotypes of mice are successfully identified, and the situation that two primers may not distinguish genotypes when large fragments are knocked out is avoided. The invention provides the convenient, reliable and economic animal model and a good foundation for researching the biological function ofPLIN1 and the pathogenesis of obesity-related metabolic diseases.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a systemic Plin1 gene knockout animal model construction and identification method. Background technique [0002] With the rapid development of the economy and the improvement of people's living standards, obesity has become an increasingly serious global health problem. In recent decades, the global incidence of overweight and obesity has increased significantly, and it is spreading rapidly in both developed and developing countries. It is generally believed that changes in the living environment in recent years are directly related to the occurrence of obesity. Unhealthy eating habits, sedentary lifestyles, physical inactivity, sleep habits, and changes in cultural and demographic background can all contribute to obesity. However, a new foreign study found that genetic factors may play a major role in the occurrence of obesity under changes in the living environment. ...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/89A01K67/027
CPCC12N15/8509C12N15/89A01K67/0276C07K14/4702C12N2800/107A01K2227/105A01K2267/0362A01K2267/03
Inventor 燕炯许祥冯晨毅刘田福陈朝阳高学坤
Owner SHANXI MEDICAL UNIV
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