Method for preparing retinitis pigmentosa non-human mammal model

Abstract

The invention relates to a method for preparing a non-human mammalian model of retinitis pigmentosa. The method comprises replacing the endogenous ctsd gene on one of the homologous chromosomes of the animal with the h-CTSD* gene; the h-CTSD* gene is a human ctsd gene with a c.262dupC site mutation. This application first discovered that the h-CTSD* gene is related to RP, and established a mouse animal model in which the h-CTSD* gene was knocked in. The invention provides a convenient, reliable and economical means for studying the relationship between CTSD mutation and retinitis pigmentosa and its pathogenic mechanism, and provides a reliable theoretical basis for the research. More importantly, the animal does not show symptoms of neurological diseases such as spasms, neuronal ceroid lipofuscin deposition, ataxia, etc., except for RP, and the pathological background is simpler, which is very suitable for the construction of RP animal models .

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...

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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, ataxia, cognitive decline, etc., and its pathological background Complex, so animals with the above mutations are difficult to use for RP mechanism research or related drug screening research

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