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Method for constructing KARS-gene-mutated mouse models and application of KARS-gene-mutated mouse models

A technology of mouse model and construction method, applied in the field of animal models, can solve the problems that it is difficult to fully reproduce the complex characteristics of Alzheimer's disease, unfavorable observation research, and high mortality

Active Publication Date: 2017-07-07
XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above-mentioned animal models are not very mature, and the construction of the model by surgery has a greater impact on the animals, the mortality rate during the operation is high, and it is difficult to survive for a long time after the operation, which is not conducive to observation and research.
The transgenic model of AD can only represent certain pathology and physiology, and it is difficult to fully reproduce the complex characteristics of Alzheimer's disease

Method used

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  • Method for constructing KARS-gene-mutated mouse models and application of KARS-gene-mutated mouse models
  • Method for constructing KARS-gene-mutated mouse models and application of KARS-gene-mutated mouse models
  • Method for constructing KARS-gene-mutated mouse models and application of KARS-gene-mutated mouse models

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Experimental program
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Embodiment Construction

[0042] The specific implementation manners of the present invention will be described below in conjunction with the accompanying drawings.

[0043] 1. Target construction:

[0044] Using CRISPR / Cas9 gene targeting technology, construct gRNA targeting the target gene, transcribe it into mRNA in vitro, and guide the Cas9 protein to cut the DNA double strand at a specific site.

[0045] (1) Gene information

[0046] Gene Bank Gene ID: 85305

[0047] (2) Design concept

[0048] Search in NCBI or ENSEMBLE based on the provided species, gene name or gene ID. Find the CDS region of the gene, that is, the coding region, analyze the corresponding genome structure, and clarify the exon part of the CDS. According to the nature of the gene itself, select candidate sites to be knocked out and determine the sites to be knocked out. After determining the knockout site, select the 23 to 250 bp exon sequence to design the sgRNA sequence, and finally select the one with less off-target as ...

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Abstract

The invention provides a method for constructing KARS-gene-mutated mouse models and application of the KARS-gene-mutated mouse models. The method for constructing the KARS-gene-mutated mouse models comprises the following steps: (1) carrying out target point construction: looking for a gene corresponding to nervous system diseases, and carrying out target point design; (2) designing Guide-RNA / Cas9mRNA, preparing plasmid, carrying out transcription, and then, injecting a transcription product into cytoplasm of a fertilized egg in a microinjection manner; (3) carrying out Founder mouse identification; (4) carrying out F1 mouse identification. According to the method for constructing the mouse models with the nervous system diseases and the application of the mouse models, provided by the invention, site-directed mutagenesis aiming at a KARS gene is carried out, so that a disease research on the model mice can be carried out under the condition of being closer to the occurrence and development of actual diseases, and a research result has a higher reference value.

Description

technical field [0001] The invention relates to a method for constructing a KARS gene point mutation mouse model, and also relates to the application of the KARS gene point mutation mouse model, which belongs to the field of animal models. Background technique [0002] Cognition is an intelligent processing process for the body to recognize and acquire knowledge, involving a series of random, psychological and social behaviors such as learning, memory, language, thinking, spirit, and emotion. Cognitive impairment refers to the pathological process in which the abnormalities in the brain's advanced intelligence processing related to the above-mentioned learning, memory, and thinking judgments lead to severe learning and memory impairments, accompanied by changes such as aphasia, apraxia, agnosia, or aphasia. [0003] Currently, the mouse models of cognitive impairment mainly use two types: vascular dementia and Alzheimer's disease. Vascular dementia is caused by acute and ch...

Claims

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Application Information

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IPC IPC(8): C12N15/85A01K67/027
CPCA01K67/0276A01K2217/075A01K2227/105A01K2267/0312A01K2267/0318C12N15/85C12N2800/80
Inventor 汪锡金杨涛余力加何龙霞宋珺王西樵
Owner XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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