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Construction and application of miRNA-34a overexpression recombinant vector

A technology for expressing vectors and recombinant vectors, applied in applications, vectors, nucleic acid vectors, etc., can solve the problems that miR-34a has not been reported yet

Inactive Publication Date: 2016-05-11
GUANGZHOU KAITUO BIOTECH DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the role of miR-34a in Alzheimer's disease or anxiety has not been reported in the prior art

Method used

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  • Construction and application of miRNA-34a overexpression recombinant vector
  • Construction and application of miRNA-34a overexpression recombinant vector
  • Construction and application of miRNA-34a overexpression recombinant vector

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Experimental program
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Embodiment Construction

[0029] The present invention will be described below through specific embodiments.

[0030] 1. Construction of miR-34a overexpression vector

[0031] 1. Sequence acquisition and primer design

[0032] Find the mouse miR-34a gene sequence (AccessionNo: EF606691) from NCBI's Nucleotide database Genbank, and then use Oligo7 software to select the partial sequences at both ends of each CDS for analysis, determine the upstream and downstream primers, and then in front of the 5' end of the upstream primer Add the protective base and BamHI restriction site sequence (CGGGATCC), and add the protective base and EcoRI restriction site sequence (CGGAATTC) to the 5' end of the downstream primer. The primers were synthesized by Shanghai Sangon Biotechnology Co., Ltd., and their sequences are shown in Table 1:

[0033] Table 1. miR-34a gene amplification primers

[0034]

[0035] 2. Obtaining template DNA

[0036] Culture Ni3T3 cells, and use Blood&CellCultureDNAMiniKit (QIAGEN) to ex...

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Abstract

The invention relates to construction and application of a miRNA-34a overexpression recombinant vector. Through constructing miR-34a overexpression adeno-associated virus recombinant vector and over-expressing miR-34a in hippocampus of mouse, the influence of miR-34a on anxiety disorder behavior can be determined. The research results show that the recombinant vector is successfully and efficiently expressed in overall animal level, the overexpression of miR-34a can cause anxiety disorder behavior, and thus miR-34a can be taken as the important candidate target for treating anxiety disorder behavior. The invention provides a novel technical means for treating diseases, which are caused by abnormal expression of miR-34A, such as anxiety disorder of Alzheimer's disease.

Description

technical field [0001] The invention relates to the construction and application of miRNA-34a overexpression recombinant vector, especially the application in the development of therapeutic drugs for Alzheimer's disease. Background technique [0002] MicroRNA is a kind of single-stranded short-sequence small RNA with a length of about 22-25 nucleotides. It does not encode protein, but it can use incomplete base pairing with the 2nd to 8th nucleotides at its 5' end Binding to the 3'UTR (3' untranslated region) of homologous mRNA, by inhibiting the expression of mRNA or inducing its degradation, it negatively regulates mRNA and affects the production of related proteins. microRNA-34a (miR-34a) is a kind of microRNA, which is widely expressed in brain tissue, myocardium, lung, liver, prostate and other tissues of the whole body. The precursor sequence of micRNA34a is shown in sequence 1, and the mature micRNA34a sequence is shown As shown in sequence 2, the precursor sequence ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/864C12N15/66A01K67/027A61K49/00
CPCA01K2267/0312A61K49/0008C12N15/66C12N15/86C12N2750/14143C12N2800/107
Inventor 罗晓斌杨细飞罗焕敏肖玲王学海徐子胜
Owner GUANGZHOU KAITUO BIOTECH DEV CO LTD