Method for preparing retinitis pigmentosa non-human mammal model

A non-human mammal and retinal pigment technology, applied in the fields of molecular biology and biomedicine, can solve problems such as complex pathological background, difficulty in screening and research of animals, and cognitive decline, and achieve the effect of simple pathological background

Active Publication Date: 2020-11-24
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] It can be seen that although some mutations of CTSD have been reported in the prior art to cause RP, they are often accompanied by a variety of neurological symptoms, such as NCL, at

Method used

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  • Method for preparing retinitis pigmentosa non-human mammal model
  • Method for preparing retinitis pigmentosa non-human mammal model
  • Method for preparing retinitis pigmentosa non-human mammal model

Examples

Experimental program
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Embodiment

[0061] This example provides the construction and identification process of the h-CTSD*-KI mouse model.

[0062] 1. Materials

[0063] The mouse strain used in this example is: C57BL / 6J, and the surrogate mother mouse strain is C57BL / 6J, which was purchased from Shanghai Slack Experimental Animal Co., Ltd., and the mice were randomly divided into a control group and an experimental group.

[0064] 2. Method

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Abstract

The invention relates to a method for preparing a retinitis pigmentosa non-human mammal model. The method comprises the following steps of replacing an endogenous ctsd gene on one of homologous chromosomes of an animal with an h-CTSD * gene, wherein the h-CTSD * gene is a humanized ctsd gene which is subjected to c.262dupC site variation. The h-CTSD * gene is discovered to be related to RP for thefirst time, and an h-CTSD * gene knocked-in mouse animal model is established. The invention provides a convenient, reliable and economic means for studying the relationship between CTSD mutation andretinitis pigmentosa and the pathogenesis of CTSD mutation and retinitis pigmentosa, and provides a reliable theoretical basis for studying CTSD mutation and retinitis pigmentosa. More importantly, besides RP, the animal does not show neurological disease symptoms such as spasm, neuron wax-like lipofuscin deposition and ataxia, the pathological background is purer, and the method is particularlysuitable for construction of an RP animal model.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and biomedicine, in particular to a method for preparing a non-human mammalian model of retinitis pigmentosa. Background technique [0002] CRISPR / Cas9 technology is a new gene editing technology developed in recent years. It is developed from the adaptive immune defense formed by archaea and bacteria under long-term selective pressure, which can effectively resist the invasion of foreign DNA. The immune system consists of three components: Cas9 protein, CRISPR RNAs (crRNAs) and trans-activating crRNAs (tracrRNA). In the process of immune defense, guide crRNA guides Cas9 protein to target foreign genes and cut them. Accordingly, people have modified crRNA-tracrRNA in vitro to obtain a sgRNA that can recognize the NGG PAM (protosp aceradjacentmotif) sequence and the target gene. At the same time, it can bind to the Cas9 protein and guide it to cut at 3 to 8 bp upstream of the PAM to form...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/89C12N15/90A61D19/04A01K67/027
CPCA01K67/0276A01K67/0278A01K2207/15A01K2217/072A01K2217/075A01K2227/105A01K2267/0306A61D19/04C12N15/113C12N15/89C12N15/907C12N2310/20
Inventor 万晓玲孙晓东
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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