Construction and identification of a systemic plin1 gene knockout animal model

Abstract

The invention belongs to the field of biotechnology and provides a systemic Plin1 gene knockout animal model construction and identification method thereof. The steps of the construction method are: designing sgRNA sequences according to the sequence before and after exon 2 of the Plin1 gene, constructing a Plin1 gene knockout vector, and performing in vitro transcription to obtain sgRNA; mixing sgRNA and Cas9 protein and then microinjecting it into mouse fertilized eggs to obtain whole body Sexual Plin1 knockout mice. The beneficial effect of this project is that a pair of sgRNA sequences with the highest scores were screened out, which ensured the success rate of the project; a large fragment of exon 2 of the Plin1 gene was knocked out, and the knockout efficiency was improved; the knockout region was designed 3 The two primers can successfully identify various genotypes of mice, avoiding the situation that two primers may not be able to distinguish genotypes when large fragments are knocked out. It provides a convenient, reliable and economical animal model and a good foundation for studying the biological function of PLIN1 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 the construction of a systemic Plin1 gene knockout animal model and an identification method thereof. Background technique [0002] With the rapid economic development 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 rapidly spreading in both developed and developing countries. It is generally believed that changes in living environment in recent years are directly related to the occurrence of obesity. Unhealthy eating habits, sedentary lifestyles, lack of physical activity, sleep habits, and changes in cultural and demographic backgrounds can all induce obesity. However, in a new foreign study, it was found that genetic factors may play a major role in the occurrence of obesity under the change of li...

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Problems solved by technology

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

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