Vector for lowering expression of cell-specific ndrg2 gene

A technology of gene expression and transgene carrier, which is applied in the field of cell-specific ndrg2 gene expression down-regulation carrier, which can solve the problems of non-tissue cell specificity and achieve obvious effect of down-regulation

Inactive Publication Date: 2011-06-15
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

[0013] Currently, RNAi transgene vectors used in RNAi transgenic animals to down-regulate the expression of target genes use a polymerase III-dependent promoter (CMV) upstream of the miRNA junction site (miR flanking region). The interfering sequence that initiates the access of the downstream miRNA junction site in the cell, so the down-regulation of the target gene expression is not tissue-cell specific

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  • Vector for lowering expression of cell-specific ndrg2 gene
  • Vector for lowering expression of cell-specific ndrg2 gene
  • Vector for lowering expression of cell-specific ndrg2 gene

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Embodiment Construction

[0042] MicroRNA (miRNA) is a kind of eukaryotic endogenous small molecule single-stranded RNA, usually 18-25 bp in length, which can cause the degradation of target mRNA or inhibit its translation through specific base pairing with target mRNA, thereby affecting Post-transcriptional expression regulation of genes is an important molecular tool in RNA interference. In recent years, hundreds of miRNAs have been discovered in animal cells and plant tissues. These small molecular regulatory RNAs are matured by cutting out the 60-200bp precursor with a hairpin structure.

[0043] In animal cells, the primary product of miRNA gene transcription (pri-miRMA) is quickly processed into miRNA precursor (pre-miRNA) by ribonuclease III Drosha (RNaseIII Drosha), and then transported from the nucleus to the cytoplasm, through another An RNaseIII Dicer recognizes spliced ​​mature miRNAs. These mature miRNAs and other proteins together form the RISC complex (RNA-induced silencing complex), wh...

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Abstract

The invention discloses a vector for lowering expression of a cell-specific ndrg2 gene, which comprises an RNAi transgene vector, wherein the RNAi transgene vector is connected with a preproinsulin promoter; the downstream of the preproinsulin promoter is inserted with an miR access site; and the miR access site is connected with an miRNA sequence aiming at lowering the ndrg2 gene. The constructedRNAi transgene vector adopts an RNA polymerase II relied preproinsulin promoter, and a specific pancreatic islet beta cell of the preproinsulin promoter initiates the downstream miRNA sequence, initiates an RNA interference aiming at the ndrg2 gene and lowers the expression of the ndrg2 gene. The vector is applied to construction of animal models and researches on ndrg2 gene functions of the pancreatic islet beta cell.

Description

technical field [0001] The invention belongs to the technical field of gene function related to type II diabetes, relates to the gene function research of type II diabetes related gene ndrg2, and particularly relates to a carrier for down-regulating the expression of cell-specific ndrg2 gene. Background technique [0002] ndrg2 and its specific expression in tissue cells [0003] The human ndrg2 gene (N-Myc Downsteam-Regulated Gene) is a new gene with low expression in tumor cells discovered and cloned by Deng Yanchun et al. (Deng Yanchun, Yao Libo, Liu Xinping. A new gene containing an ACP-like domain Discovery Biochemistry and Biophysics Progress. 2001, 28 (1): 72-76), because this gene has a relatively similar structure with the N-myc down-stream regulated gene ndrg1 (N-myc down-stream regulated gene 1) that has been discovered High homology, so it was named ndrg2. The full-length cDNA of the gene is 2024bp, encoding 357 amino acids, located at 14q12.1, with 16 exons an...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/85
Inventor 刘新平吴有盛药立波
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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