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Methyl CpG binding protein 2-based (MECP2-based) method for injecting virus into specific brain area of animal without therapeutic aim to perform gene editing

A technology of gene editing and viruses, which is applied in the field of constructing non-human primate ASD disease models, can solve the problems of unsolvable chimerism of embryo transgenic technology, difficult research needs, high mortality rate of transgenic monkeys, etc., to avoid embryo transgenic technology Issues of chimerism, avoidance of genetic differences, effects of addressing research needs

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

[0004] At present, there are only a few embryonic ASD macaque models for macaques, but (1) the genetic background of macaques varies greatly among individuals, and the problem of mosaicism in embryo transgenic technology cannot be solved, so it is still difficult for these macaques based on embryo transgenes to help us To observe and study the etiology and pathogenesis of ASD
(2) The natural sexual maturity time of macaques is 5-6 years, and the gestation period takes 6 months. They are singleton animals. Therefore, it is difficult to achieve genetically manipulated monkeys that are homozygous by mating transgenic macaques at this stage.
(3) Transgenic monkeys based on embryo transgenic technology have high mortality rate, low success rate, poor stability of transgenic gene retention, and extremely long onset time. At present, it is difficult to establish a stable transgenic monkey model
Even if it is successfully established, it is difficult for this monkey model to reproduce in large numbers and the maintenance cost is extremely high, so it is difficult to meet the urgent research needs at this stage

Method used

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  • Methyl CpG binding protein 2-based (MECP2-based) method for injecting virus into specific brain area of animal without therapeutic aim to perform gene editing
  • Methyl CpG binding protein 2-based (MECP2-based) method for injecting virus into specific brain area of animal without therapeutic aim to perform gene editing
  • Methyl CpG binding protein 2-based (MECP2-based) method for injecting virus into specific brain area of animal without therapeutic aim to perform gene editing

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Embodiment Construction

[0013] At present, the injection of animal models for this disease should focus on rodents, mainly mice. Rodents are far from human relatives, lack a correspondingly developed cerebral cortex, and have simple social behaviors. Their brain development process is similar to that of humans. Also very different. (Johnson, Wang et al.2015), non-human primates, especially macaques, have a genetic similarity with humans as high as 93.5%, and are very similar to humans in terms of physiological structure, developmental process, brain function and behavioral performance . More importantly, the macaque is a naturally social animal, which has obvious advantages over other animals for the study of diseases with typical social disorders such as ASD.

[0014] Therefore, the embodiment of the present invention proposes a method of directly injecting virus into a specific brain region of a non-human primate for gene editing, and the rhesus monkey is used as an example for illustration below....

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Abstract

The invention discloses a methyl CpG binding protein 2-based (MECP2-based) method for injecting a virus into a specific brain area of an animal without a therapeutic aim to perform gene editing. A cerebral stereotaxis injection pump is utilized to directly inject the virus into the specific brain area of a non-human primate; the injection volume is that 1 [mu]L of virus is injected per 1m<3>; andthe titer of the injected virus is 10*10<13> vg / mL. According to the method, the virus is directly injected into the specific brain area of the non-human primate to perform gene editing, and thus, a transgenic monkey model with the disease phenotype can be obtained in a short time, and the urgent research demand of an autism spectrum disorder (ASD) disease at the present stage is 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 non-therapeutic method based on the MECP2 gene that injects viruses into specific brain regions of non-human primates for gene editing to construct non-human primates. Methods for animal-like ASD disease models. Background technique [0002] Autism Spectrum Disorder (ASD) is characterized by autism-like behavior, loss of motor control, irregular breathing, and skeletal problems. Adolescents with ASD cannot participate in the normal education process, which has a huge impact on their normal integration into society and their social function as adults. At present, there is no effective treatment for ASD diseases, so ASD has a serious negative impact on family happiness and harmonious development of society, and has been widely valued by all walks of life. [0003] In recent years, the prevalence of ADS has been rising and has be...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N7/01C12N15/864C12N15/113C12N15/90A01K67/027
CPCC12N15/8509C12N7/00C12N15/86C12N15/113C12N15/907A01K67/0276C12N2310/20C12N2750/14121C12N2750/14143A01K2207/15A01K2217/075A01K2227/106A01K2267/0356
Inventor 吴诗昊李霄程田林秦冬冬吴晶胡新天仇子龙
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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