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AAV9-CIP virus of expressed neurodegenerative disease protection peptide and preparation method of AAV9-CIP virus

A AAV9-CIP, neurodegenerative technology, applied in the medical field, can solve the problems of not being able to pass the blood-brain barrier and difficult clinical transformation

Active Publication Date: 2017-02-01
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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Problems solved by technology

However, because the molecular weight of CIP is too large (13.8kDa), it cannot pass through the blood-brain barrier, so it is difficult to be used for clinical transformation

Method used

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  • AAV9-CIP virus of expressed neurodegenerative disease protection peptide and preparation method of AAV9-CIP virus
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  • AAV9-CIP virus of expressed neurodegenerative disease protection peptide and preparation method of AAV9-CIP virus

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[0076] Before setting forth the method of the present invention, the following abbreviations are explained and described:

[0077] Myc-CIP primer: The 5' end of the primer is designed to introduce a restriction endonuclease EcoRI restriction site and the specific sequence of c-Myc-tag. The introduction of the EcoRI restriction site is used for subsequent Myc-CIP gene cloning In other vectors, Myc sequence and CIP maintain the same reading frame, and 10 peptides of Myc will be added to the N-terminus of CIP, so that the expressed fusion Myc-CIP fusion protein can be recognized by c-Myc monoclonal antibody, available In the follow-up work, the identification and recognition of CIP protein in vivo, and because the Myc peptide is short, it is often used to help other protein recognition without affecting the function of other proteins.

[0078] Myc-CIP primer: The 5' end of the primer is designed with the specific sequence of c-Myc-tag, so that the expressed CIP protein can be rec...

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Abstract

The invention discloses a preparation method of an AAV9-CIP virus of expressed neurodegenerative disease protection peptide. The preparation method comprises the following steps that 1, a Myc-CIP primer is designed, and Myc-CIP gene segments are obtained; 2, plasmid pCR2.1-Myc-CIP is obtained; 3, plasmid pEGFP-C2-Myc-CIP is obtained; 4, plasmid pAAV-hSynapsin-GFP-Myc-CIP is obtained; 5, the AAV9-CIP virus is obtained. The invention aims at providing the AAV9-CIP virus, capable of being specifically expressed in central nervous system neuron, of neurodegenerative disease protection peptide. Due to the fact that a neurotoxic compound CDK5 / p25 generated in the lesion process of the nervous system neurodegenerative disease can be specifically inhibited by CIP, the CIP virus capable of being used for targeted intervention treatment of the nervous system neurodegenerative disease, such as the Alzheimer's disease and Parkinson's disease is established.

Description

technical field [0001] The invention relates to the medical field, in particular to an AAV9-CIP virus expressing a neurodegenerative disease protection peptide and a preparation method thereof. Background technique [0002] Current human and animal experiments have proved that the high activity of CDK5 / p25 plays an important pathological role in neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, lateral sclerosis, and stroke. CIP is a fragment of 125 amino acid residues at the end of p25. In vitro neuron protection experiments and hybridization studies between CIP transgenic mice and Alzheimer's disease mouse models found that CIP peptides can specifically inhibit the toxicity of CDK5 / p25 and play a role in neurons. Protective effects. However, because the molecular weight of CIP is too large (13.8kDa), it cannot pass through the blood-brain barrier, so it is difficult to be used for clinical transformation. Contents of the invention [0003] Th...

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Application Information

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IPC IPC(8): C12N15/864C12N7/01
CPCC07K14/4748C12N7/00C12N15/86C12N2750/14121C12N2750/14143
Inventor 胡亚芳黄建欧潘速跃许苗菁贺荣霓姬仲周晓何勇宋萍萍顾勇谢作善李伟刘光辉
Owner NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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