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Construction method of Amy1 gene knockout mouse animal model and application

A technology of gene knockout mice and animal models, applied in the field of biology

Active Publication Date: 2020-03-17
SHANGHAI JIAO TONG UNIV
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  • Description
  • Claims
  • Application Information

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However, there is no report on the construction method of the Amy1 knockout mouse animal model

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  • Construction method of Amy1 gene knockout mouse animal model and application
  • Construction method of Amy1 gene knockout mouse animal model and application
  • Construction method of Amy1 gene knockout mouse animal model and application

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Embodiment Construction

[0043] The present invention will be described in detail below in conjunction with examples. The following examples will help those skilled in the art to further understand the present invention, but do not limit the present invention in any form. It should be noted that those skilled in the art can make some adjustments and improvements without departing from the concept of the present invention. These all belong to the protection scope of the present invention.

[0044] 1. The construction scheme of Amy1 gene knockout mouse, the schematic diagram is as follows figure 1 shown;

[0045] (1) Construction of specific target sgRNA1 and sgRNA4 for Amy1 gene. DNA was linearized and purified and transcribed into mRNA in vitro. The sgRNA1 sequence of the Amy1 is shown in SEQ ID NO.1; the sgRNA4 sequence of the Amy1 is shown in SEQ ID NO.2;

[0046] The gRNA sequence information and transcript concentration are shown in Table 1:

[0047] Table 1

[0048] gRNA name gRN...

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Abstract

The invention discloses a construction method of an Amy1 gene knockout mouse animal model and application. An Amy1 gene knockout mouse model is constructed based on a CRISPR / Cas9 gene knockout technology. The construction method comprises the following steps: step 1, designing sgRNA and Cas9 RNA, performing in-vitro transcription to obtain mRNA, and performing microinjection of the active sgRNA and Cas9 RNA on fertilized eggs of a mouse to obtain an Amy1 gene knockout mouse; and step 2, identifying the Amy1 gene knockout mouse animal model. The method has advantages: the CRISPR / Cas9 gene knockout technology is used and the mouse animal model with the Amy1 gene knocked out is established for the first time; and a convenient and reliable animal model is provided for researching the relationbetween the Amy1 gene and related diseases.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a construction method and application of an Amy1 knockout mouse animal model based on CRISPR / Cas9 technology. Background technique [0002] The mouse Amy1 gene is located on the reverse strand of chromosome 3, with a full length of 24190bp, which encodes salivary gland amylase. Salivary amylase is 511 amino acids in length, mainly involved in the metabolism of carbohydrates, and plays the role of hydrolyzing starch into maltose and dextran. Salivary amylase acts on the α-1,4 glycosidic bond, and can decompose the larger molecular weight and insoluble starch molecules into soluble small particles (including starch dextrin, red dextrin and colorless dextrin), which are then absorbed in the stomach Gastric acid inactivation. The substrate of salivary amylase—starch and the product of its metabolic process—short-chain glucose can partially protect salivary amylase from bein...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/90C12N15/85C12N15/56A01K67/027
CPCC12N15/907C12N15/8509C12N9/2411A01K67/0276A01K2217/075A01K2227/105A01K2267/0362C12N2800/107
Inventor 隋中泉徐义娟高海德李紫君马梦婷温雅迪
Owner SHANGHAI JIAO TONG UNIV
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