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Sequence of hepatitis B virus (HBV) specific microRNA like siRNA (msiRNA) and application thereof

A hepatitis B virus, specific technology, applied in the field of genetic engineering, can solve problems such as immature solutions

Inactive Publication Date: 2014-09-24
SHIYAN TAIHE HOSPITAL
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Problems solved by technology

[0003] Combination therapy of antiviral and immune regulation is a recognized research direction, but the program is not yet mature

Method used

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  • Sequence of hepatitis B virus (HBV) specific microRNA like siRNA (msiRNA) and application thereof
  • Sequence of hepatitis B virus (HBV) specific microRNA like siRNA (msiRNA) and application thereof
  • Sequence of hepatitis B virus (HBV) specific microRNA like siRNA (msiRNA) and application thereof

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Embodiment Construction

[0016] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0017] figure 1 It is the secondary structure of microRNA-like siRNA specific for hepatitis B virus.

[0018] (1) msiHBs inhibited HBV antigen expression in Huh7 cells.

[0019] HBVS gene-specific msiRNA and reporter HBV replication plasmid pHY106+wta were co-transfected into Huh7 cells, and the supernatant was collected at 72 hours, and the expression of HBsAg was detected by ELISA. As a result, msiHBs significantly inhibited HBsAg expression ( figure 2 ).

[0020] (2) msiHBs inhibited HBV antigen expression in HepG2 cells.

[0021] HBVS gene-specific msiRNA and reporter HBV replication plasmid pHY106+wta were co-transfected into HepG2 cells, and the supernatant was collected after 72 hours, and the expressions of HBsAg and HBeAg were detected by ELISA. As a result, msiHBs significantly inhibited the expression of HBsAg and HBeAg ( i...

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Abstract

The invention provides a sequence of microRNA like siRNA (msiRNA) aiming at a hepatitis B virus (HBV) S gene and a secondary structure of msiRNA. The sequence is a nucleotide sequence of msiHBs. After co-transfecting HepG2 and Huh7 cells, the msiRNA and a reporter HBV replicating plasmid pHY106+wta can inhibit expression of HBsAg and HBeAg. After transfecting the HepG2.2.15 cell, the msiHBs can also effectively inhibit the expression levels of HBsAg and HBeAg and the load of the HBV in the supernatant. Besides, the msiRNA also has the immunostimulatory activity and can activate innate immunity via a Toll like receptor pathway. Therefore, the msiRNA has double activities of inhibition of HBV gene expression as well as replication and immunostimulation and can be applied to preparation of drugs for treating HBV chronic infection.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a microRNA-like siRNA sequence specific to hepatitis B virus and its application. Background technique [0002] Hepatitis B virus (HBV) infection is one of the important causes of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. There are about 93 million chronic HBV-infected people in my country, and research on the prevention and treatment of hepatitis B has always been an important field of medical and health research. So far, there is still a lack of long-term effective antiviral drugs for HBV infection. Although the treatment of interferon (IFN-α) and nucleoside (acid) analogs can achieve certain curative effect, there are still problems such as low sustained response rate, certain side effects, and easy drug resistance after long-term application. Studies have shown that in patients with chronic hepatitis B, the immune system is not perfect, the ...

Claims

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Application Information

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IPC IPC(8): C12N15/113A61P31/20
Inventor 孟忠吉张永红陈悦
Owner SHIYAN TAIHE HOSPITAL
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