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Method for improving social intercourse disorders of autism

A technology for patients with social disorders and autism, applied in the biological field, it can solve the problems of not finding specific brain areas and the inability to provide patients with methods, and achieve the effect of improving social behavior disorders

Inactive Publication Date: 2020-05-15
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] For the treatment of autism mouse models, it is still in the stage of mutant wild-type hybridization or the change of MeCP2 gene level in the whole brain, and no specific brain area related to autism has been found, nor is it specific in this brain area. Knock down the MeCP2 gene to observe its changes, and these methods are not available to patients

Method used

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  • Method for improving social intercourse disorders of autism
  • Method for improving social intercourse disorders of autism
  • Method for improving social intercourse disorders of autism

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Example 1: Comparison of three-compartment behavior between WT and MECP2-TG autistic mice

[0045] 1) Three-compartment experiments were performed on WT mice and MECP2-TG autistic mice (15 weeks). See attached Figure 5 a, Close a strange mouse in the cage of the leftmost box, record the communication between WT mice and MECP2-TG autistic mice and the strange mouse, and record their trajectories in 10 minutes . At the same time, use the recorded data to make a time distribution heat map in matlab.

[0046] attached Figure 5 In a, the contact area is between the gray box and the white dashed box (cage area) inside, and the distance between the two is 4.5 cm. If there are juvenile mice (strange mice) in cages in the area, this area is called a social zone (SocialZone, abbreviated as SZ, and the cells that carry out relevant and intense activities in the SZ area are called SZ cells). The body is an SZ box. If there is no juvenile mouse in the cage in the area, the a...

Embodiment 2

[0060] Example 2: Construction process of AAV-Cas9-sgRNA1 / 2 for specific knockdown of hMECP2 gene

[0061]1) Import the full length of the human hMeCP2 gene into the http: / / www.e-crisp.org / E-CRISP / website, search for a suitable PAM site and a suitable sgRNA, and obtain more than two sgRNAs. The experiment of the present invention Two sgRNAs, 5'-CCTGGGGCTCAGGGGGGCTGGTGGGGT-3' (sgRNA1, shown in SEQ ID NO: 1) and 5'-ACTCTGAGTGGTGGTGATGGTGGTGG-3' (shown in SEQ ID NO: 2), are used, as shown in the attached image 3 .

[0062] 2) Two sgRNAs were synthesized according to the principles of primer design, and constructed into the plasmid pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA; U6::BsaI-sgRNA (purchased from addgene) after annealing. The backbone of the vector is modified, the CMV fragment is excised and inserted into the synapsin promoter, and the HA tag is excised and inserted into the Flag tag. Transformation into pX601-AAV-hMECP2::NLS-SaCas9-NLS-3xflag-bGHpA; U6::BsaI-sgRNA1 / 2 ...

Embodiment 3

[0063] Example 3: The process of specifically knocking down the human hMeCP2 gene in the hippocampal CA1 region of MECP2-TG mice

[0064] During the experiment, on the one hand, it is necessary to perform Cas9-mediated gene knockdown on MECP2-TG mice (administering AAV-flag-SaCas9-SghMECP1 / 2, that is, containing the plasmid pX601-AAV-hMECP2::NLS-SaCas9-NLS -3xflag-bGHpA; U6:: AAV virus of BsaI-sgRNA1 / 2) Preparation of treatment group mice (Rescue): On the other hand, it is necessary to perform functional fluorescence detection on neurons in the hippocampus CA1 area of ​​the treatment group mice. Neurons in the CA1 region were infected with another AAV virus (AAV2 / 5-DIO-GCaMP6s, purchased from Vector biolabs).

[0065] 1) AAV-flag-SaCas9-SghMECP2 and AAV2 / 5-DIO-GCaMP6s were mixed 1:1 (total volume 1 μl) and loaded into the glass electrode on the stereotaxic instrument.

[0066] 2) Use a stereotaxic instrument to find the CA1 area of ​​the mouse (1.9mm backward from bregma, 1.4...

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Abstract

The invention relates to the technical field of biology, in particular to a method for improving social intercourse disorders of autism, and provides a method which can improve social intercourse disorders of an autism mouse model. According to the method, a specific knock-down vector specific to human-derived methyl CpG binding protein 2 (hMECP2) is constructed, and the specific knock-down vectoris injected into a hippocampal cornu ammonis 1 (CA1) region of the MECP2-transgene (MECP2-TG) autism mouse model by a stereotactic injection method, and thereby, the social intercourse disorders of the autism mouse model are improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for improving the social impairment of autistic patients. Background technique [0002] Autism, also known as autism, is a subtype of pervasive developmental disorder, more common in males, with onset in infancy, mainly manifested in different degrees of language development disorders, social communication behavior disorders and repetitive stereotyped behaviors . The pathogenesis of autism is complex, and the main factors include genetics, perinatal factors, immune system abnormalities, and neuroendocrine abnormalities. According to data from the National Institute of Mental Health (NIMH), the prevalence of autism in the United States is between 1‰ and 2‰. In 2010, related reports from some regions in China showed that the prevalence of autism in Guangdong was 0.67%, and it was as high as 1.32% in Shenzhen. The social impairment of autistic patients is closely related to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P25/00A61K49/00
CPCA61K48/005A61P25/00A61K49/0008
Inventor 王晓群仇子龙吴倩孙乐
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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