shRNA molecule capable of inhibiting expression of human AEBP1 gene

A gene expression and molecular technology, applied in the field of shRNA molecules that inhibit the expression of human AEBP1 gene, to achieve the effect of reducing expression

Inactive Publication Date: 2016-09-21
刘媛
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However, there is no shRNA molecule tha

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  • shRNA molecule capable of inhibiting expression of human AEBP1 gene

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Embodiment Construction

[0020] The technical solutions of the present invention will be clearly and completely described below in conjunction with specific embodiments.

[0021] In the present invention, the shRNA molecular synthesis method for inhibiting human AEBP1 gene is as follows:

[0022] According to the human AEBP1 gene in Genbank, gene sequence number: NM_001129.4; design 4 shRNA molecules, and synthesize two complementary single-stranded shRNA molecules according to the shRNA sequence, namely shRNA1 and shRNA2. These two shRNA molecules are annealed to form double strands Finally, connect to the lentiviral vector to construct 4 lentiviral interfering RNA plasmids.

[0023] In order to test the inhibitory effect of the synthetic shRNA molecule on the expression of human AEBP1 gene, the following method was used to transfect the shRNA plasmid into the Huh7 cell line containing AEBP1: the Huh7 cells expressing AEBP1 were inoculated into a 24-well plate, 1.5×10 5 Cells / well density, culture c...

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Abstract

The invention discloses a shRNA molecule capable of inhibiting expression of the human AEBP1 gene. According to the human AEBP1 gene (with a serial number of NM_001129.4) in Genbank, four shRNA molecules are designed and are synthesized targeted at the sequence of shRNA; two synthesized complementary single-stranded shRNA molecules are subjected to annealing to form a double-stranded shRNA molecule; and the double-stranded shRNA molecule is linked to a lentivirus vector to construct four lentivirus interference RNA plasmids. The shRNA plasmids are used for transfection of a Huh7 cell line containing AEBP1, and relative quantitative analysis is carried out on the AEBP1 gene through real-time quantitative PCR so as to evaluate inhibition effect on the expression of the AEBP1 gene. Results of relative quantitative analysis show that AEBP1-shRNA1 can inhibit transcription of 82.3% of AEBP1 and AEBP1-shRNA2 can inhibit transcription of 75.7% of AEBP1.

Description

technical field [0001] The invention relates to the design, synthesis and evaluation method of the shRNA molecule capable of significantly inhibiting the adipocyte enhancer-binding protein 1 (Adipocyte Enhancer-binding Protein 1, AEBP1) gene and the inhibitory effect on liver cells. Background technique [0002] Adipocyte enhancer-binding protein 1 (AEBP1) is a transcriptional repressor with a molecular weight of 82kDa. It is highly expressed in adipose tissue and participates in adipogenesis in preadipocytes. In addition, the expression levels of AEBP1 in liver, lung, spleen, brain and primary macrophages were also higher. [0003] Studies have shown that AEBP1 is involved in the cholesterol balance of mouse macrophages, the formation of foam cells, and the inflammatory response of macrophages and liver cells. High expression of AEBP1 promotes the release of inflammatory factors in mice. The specific mechanism is that AEBP1 promotes the release of inflammatory factors suc...

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Application Information

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IPC IPC(8): C12N15/113
CPCC12N15/113C12N2310/10
Inventor 刘媛
Owner 刘媛
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