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Method for establishing fragile X-syndrome non-human primate model on basis of CRISPR gene knockout technology

A primate, gene knockout technology, applied in the field of biomedicine, can solve problems such as no primate model, and achieve the effect of reducing risk

Inactive Publication Date: 2014-03-19
SUZHOU TONGSHAN BIO TECH
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AI Technical Summary

Problems solved by technology

[0006] In the study of FXS disease, so far there are no reports of primate models at home and abroad

Method used

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Embodiment 1

[0031] A method for establishing a fragile X syndrome primate model based on CRISPR gene knockout technology, comprising the following steps:

[0032] (1) Establishment of FMR1 gene knockout cynomolgus monkey model;

[0033] Proceed as follows:

[0034] 1) Construction of CRISPR targeted modification gene carrier;

[0035] 2) Ovulation induction and in vitro fertilization in cynomolgus monkeys;

[0036] 3) Microinjection of cynomolgus monkey fertilized eggs;

[0037] 4) In vitro culture of fertilized eggs, implantation into recipients and cultivation of targeted gene-modified animals;

[0038] (2) Identification of cynomolgus monkey animal model and related functional analysis;

[0039] Proceed as follows:

[0040] 1) DNA sequencing of the CRISPR target site to identify the genotype of the cynomolgus monkey animal model;

[0041] 2) Using real-time quantitative PCR (real-time PCR) to detect the decrease of FMR1 gene mRNA level in cynomolgus monkey cells;

[0042] 3) Det...

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Abstract

The invention discloses a method for establishing a fragile X-syndrome non-human primate model on the basis of a CRISPR gene knockout technology. The method comprises the following steps: (1) establishing a FMR1 gene knockout machin model; (2) carrying out identification and related functional analysis on the machin model; (3) carrying out tests on the nerve characteristics and learning and memorizing ability of the machin model. The method utilizes a CRISPR gene knockout technology to establish a fragile X-syndrome non-human primate model. The model fills the blank of non-human primate model, can effectively stimulate the pathological process of human diseases, can be used as an optimum animal model for researching human diseases, can effectively predict the effect of novel vaccine, novel drug or novel diagnostic reagent in clinical applications, and thus greatly reduces the risk of novel drug development.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for establishing a primate model of fragile X syndrome based on CRISPR gene knockout technology. Background technique [0002] Fragile X syndrome (FXS) is the most common genetic mental retardation linked to the X chromosome, affecting approximately 1 / 3600 males and 1 / 4000-6000 females. The cause is FMR1 CGG high-copy dynamic mutation occurs in the gene, resulting in FMR1 Gene methylation (DNA methylation) and gene silencing (gene silencing), mental retardation protein (FMRP) is not expressed or the expression level is reduced. [0003] Knowledge about the pathogenesis of FXS comes primarily from studies in FMR1 knockout mouse models. The FXS mouse model exhibits some clinical symptoms similar to human patients, such as: tremor, motor ataxia, attention deficit, intellectual disability. At the molecular level, due to the reduction or absence of FMRP expression, the expressi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85A01K67/027
Inventor 薛志刚范国平林卿
Owner SUZHOU TONGSHAN BIO TECH
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