Non-therapeutic-purpose gene editing method based on PINK1 and PARK7 through injection of virus into specific brain regions of animals

A technology of gene editing and viruses, applied in the direction of single-stranded DNA viruses, other methods of inserting foreign genetic materials, plant gene improvement, etc., can solve the problems of not being an etiological model, the cause of disease, and the powerlessness of disease mechanism, so as to avoid embryonic The problem of mosaicism in transgenic technology, long molding time, and the effect of avoiding genetic differences

Pending Publication Date: 2020-06-19
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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Problems solved by technology

[0003] Although the classic MPTP macaque model has been widely used in many aspects of PD research, it is basically powerless t

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  • Non-therapeutic-purpose gene editing method based on PINK1 and PARK7 through injection of virus into specific brain regions of animals
  • Non-therapeutic-purpose gene editing method based on PINK1 and PARK7 through injection of virus into specific brain regions of animals
  • Non-therapeutic-purpose gene editing method based on PINK1 and PARK7 through injection of virus into specific brain regions of animals

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Embodiment Construction

[0011] Rhesus macaques have good kinship with humans, have very similar brain structures, and their genes are as high as 93% similar to human genes (Huang, Zhang and Chen 1993). In addition, macaques have advanced brain functions such as reasoning, analysis, decision-making, and memory, and can complete the most complex behavioral tasks other than humans. Compared with other animals, it is easier to perform complex and accurate behavioral tests. Based on this, we plan to use AAV virus as a vector to mutate the PINK1 and PARK7 genes of rhesus monkeys using CRISPR / CAS9 technology, and use MRI imaging as a precise guide to specifically inject the virus into the unilateral or bilateral substantia nigra of rhesus monkeys to establish the virus. Transgenic PD macaque model. Then use the recognized macaque PD behavior scale (Kurlan scale) and apomorphine-induced rotation behavior to evaluate the PD behavior symptoms of model animals, and perform immunohistochemical pathological verif...

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Abstract

The invention discloses a non-therapeutic-purpose gene editing method based on PINK1 and PARK7 through injection of a virus into specific brain regions of animals. The virus is directly injected intothe specific brain regions of non-human primates by using a brain stereotaxic injection pump, wherein the injection volume is 1 [mu]L of the injected virus per 1 m<3>, and the titer of the injected virus is 10*10<13>vg/mL. Through the method, gene editing is performed directly on the specific brain regions of the non-human primates, and transgenic monkey models with disease phenotypes can be obtained in a short time, so that urgent research needs of Parkinson's syndromes (PD) at the present stage are met.

Description

technical field [0001] The invention belongs to the field of transgenic technology and modeling of non-human primates, and specifically relates to a method for injecting viruses into non-human primates based on PINK1 and PARK7 genes (note: PARK7 and DJ-1 are the same gene) for non-therapeutic purposes. A method for gene editing of specific brain regions in animals to construct PD disease models in non-human primates. Background technique [0002] Parkinson's disease (PD) is a typical neurodegenerative disease. The etiology and mechanism of the disease are unknown, and it mainly occurs in the elderly population over the age of 65. At present, there is no cure. While causing great pain to the patients, it also brings a heavy burden to the families and society. Due to their high evolutionary similarity, the non-human primate model is currently one of the most suitable models for studying the pathogenesis of PD and developing effective treatments. [0003] Although the classic...

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Application Information

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IPC IPC(8): C12N15/864C12N15/90C12N15/54C12N15/55A01K67/027
CPCC12N15/86C12N15/907C12N9/12C12N9/80C12Y207/11001A01K67/0276C12N2750/14143C12N2800/107A01K2217/075A01K2227/106A01K2267/0318
Inventor 吴诗昊李浩李霄程田林吴晶仇子龙胡新天
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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