SiRNA for specifically inhibiting ECHS1 gene expression and application thereof

Abstract

The invention discloses siRNA for specifically inhibiting ECHS1 gene expression and application thereof. The small interfering RNA is double-stranded RNA consisting of nucleotide sequences shown in a sequence 1 of a sequence table and a sequence 2 of the sequence table. The invention provides the siRNA which can specifically and efficiently inhibit the ECHS1 gene expression and modified RNA obtained by modifying the siRNA. In vitro and in vivo experiment results show that the RNA can specifically and efficiently inhibit the ECHS1 gene expression, promote the increase of free fatty acid and the deposition of triglyceride, and show the increase of lipid droplets. The siRNA can be applied to the preparation of a nonalcoholic fatty liver disease animal model and the research on a nonalcoholic fatty liver disease pathogenesis. The siRNA has important values.

Description

technical field [0001] The invention relates to a siRNA specifically inhibiting the expression of ECHS1 gene and its application. Background technique [0002] RNA interference (RNA interference, RNAi) is an evolutionarily conserved defense mechanism against the invasion of transgenes or foreign viruses. Double-stranded RNA (dsRNA) specifically degrades the mRNA in the cell, thereby resulting in the process of effectively blocking the specific gene, which is a sequence-specific post-transcriptional gene silencing (PTGS). RNAi technology has been successfully applied to the study of gene functions in various organisms. Construct siRNA for the target gene, use RNAi technology to shut down the expression of the gene, and analyze the function of the gene according to the change of the phenotype. Gene knockout technology takes a long time to permanently shut down the expression of a certain gene, while RNAi technology can controllably shut down 10 genes within 1 week or even 2 ...

AI Technical Summary

Problems solved by technology

[0005] The molecular mechanism of the occurrence and development of NAFLD has not yet been clarified. At present, NAFLD is believed to be related to insulin resistance, fat storage disorder, and fat secretion inhibition. Among them, fat storage disorder is one of the hotspots in the study of the pathogenesis of NAFLD. Normal liver fatty acid oxidation and synthesis are in a dynamic balance , Once fatty acid oxidation disorder, fat accumulation in the liver, fatty liver occurs. Mitochondrial β-oxidation is the main oxidation form of fatty acid oxidation. This process can be divided into three stages: activation, transfer, and β-oxidation. The activated fatty acid is converted into acyl CoA The form of β-oxidation is transferred to the mitochondrial matrix for β-oxidation. β-oxidation undergoes four-step reactions, namely dehydrogenation, water addition, dehydrogenation and thiolysis, to generate a molecule of acetyl CoA and a new fatty acyl CoA with two less carbons. More Repeating the cycle above will gradually generate acetyl-CoA. Among them, the second step of adding water is catalyzed by ECHS1 (enoyl-CoA hydratase). The reduction of ECHS1 expression may lead to NAFLD, but there is no correlation between ECHS1 and non-alcoholic hydratase. The study of the mechanism of ECHS1 in the pathogenesis of NAFLD using RNAi technology is an important supplement to the pathogenesis of NAFLD.

[0006] The existing animal models of non-alcoholic fatty liver disease, such as ob / ob, db / db mice and Obese Zucker rats and other genetically derived spontaneous animal models, mostly need to be imported from abroad, are expensive, and require high feeding conditions , and these animal models cannot spontaneously reproduce non-alcoholic steatohepatitis due to the lack of leptin gene; while the non-alcoholic fatty liver disease animal model reproduced by feeding SD rats with a high-fat diet can reproduce non-alcoholic fatty liver disease steatohepatitis, but the molding speed is slow, generally takes about six months, and the stability of the model is not good

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