Construction of si RNA expression vector of tissue inhibitor of metalloproteinases (TIMPs) and application in treatment of cirrhosis

Abstract

The invention discloses the construction of a si RNA expression vector of the tissue inhibitor of metalloproteinases (TIMPs) and application in treatment of cirrhosis, which belong to the technical field of genetic engineering. In the invention, according to the design principle of siRNA, a corresponding si RNA sequence is designed by selecting nucleotide sequences of TIMP-1 and TIMP-2 coding areas respectively to perform BLAST comparison, so that isogenesis with other genes is avoided; single-stranded oligonucleotides chemically synthesized and the double stranded DNA formed by annealing in vitro are connected with a eukaryotic expression vector pGeneil-1; and enzyme digesting identification and sequence assay prove that the si RNA expression vector of the TIMP-1 and TIMP-2 is constructed successfully. The vector transfects a hepatic stellate cell (HSC) through liposome and can inhibit the expression of the TIMP-1 and the TIMP-2 obviously; cirrhosis animal model assay shows that the expression vector can reduce the collagenous fiber deposition of liver tissues considerably and is expected to be used as a novel anti-hepatic fibrosis genetic therapy in clinical treatment.

Description

Technical field: [0001] The invention belongs to the technical field of genetic engineering Background technique: [0002] Traditional gene therapy (gene therapy) is a treatment method that uses normal or wild type (wild type) genes to correct or replace disease-causing genes. In this treatment method, the target gene is introduced into the target cells, and they either integrate with the host cell chromosome to become part of the host genetic material, or do not integrate with the chromosome and are located outside the chromosome, but both It can be expressed in cells and play a role in treating diseases. At present, the concept of gene therapy has been greatly expanded. Any disease treatment method carried out at the nucleic acid level using the methods and principles of molecular biology can be called gene therapy. With the in-depth understanding of the nature of the disease and the continuous emergence of new molecular biology methods, gene therapy has made great progr...

AI Technical Summary

Problems solved by technology

It is precisely because of the increase of TIMPs that ECM cannot be degraded by MMPs in time and accumulates, leading to liver fibrosis and even liver cirrhosis

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