siRNA for treating hbv

A technology for HBV and treatment, applied in the field of genetic engineering, can solve the problems of chronic HBV patients obtaining expected results and being in a state of immune tolerance

Inactive Publication Date: 2011-01-19
SHANDONG UNIV
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Problems solved by technology

However, most of the chronic HBV patients did not get the expected results under conventional treatment
Although interferon can inhibit the replication of HBV, HB

Method used

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  • siRNA for treating hbv
  • siRNA for treating hbv
  • siRNA for treating hbv

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Experimental program
Comparison scheme
Effect test

Embodiment

[0062] 1. Synthesis of chemically synthesized siRNA:

[0063] Design siRNA for HBV X gene: According to the siRNA design principle, three pairs of siRNA were handed over to Guangzhou Ruibo Company to directly synthesize the siRNA sequence, see Table 1, and the sequence table SEQ ID NO.1-SEQ ID NO.3.

[0064] 2. In vitro transcription of 3p-siRNA:

[0065] (1) The target and sequence are completely consistent with chemically synthesized siRNA.

[0066] (2) Synthesize single-stranded DNA corresponding to the HBx-siRNA sequence with T7promoter, see Table 2.

[0067] (3) Preparation of double-stranded oligo DNA: the synthesized single-stranded oligo DNA was dissolved in DEPC-treated water to prepare a 100 pmol / μl DNA solution.

[0068] ① Prepare Oligo DNA annealing reaction solution according to the components shown in Table 3.

[0069] ②Place the Oligo DNA annealing reaction solution above on a PCR amplification instrument, treat it at 95°C for 2 minutes, cool it to 25°C for 4...

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Abstract

The invention discloses siRNA for treating HBV, which has a sequence shown in SEQ ID NO.1 or SEQ ID NO.2 or SEQ ID NO.3. The siRNA can be artificially synthesized or synthesized through in vivo transcription. After the siRNA transfects HepG2.2.15 cell for 24 hours and 48 hours, the siRNA can reduce the expression of HBx gene, the content of HBsAg and HBeAg in the cell and supernate is also obviously reduced, and the effect of 3p-siRNA is more obvious than that of chemically synthesized siRNA. The siRNA can be applied to medicaments for resisting the HBV.

Description

technical field [0001] The invention relates to a siRNA used for treating HBV, which belongs to the field of genetic engineering. Background technique [0002] At present, there are about 420 million HBV carriers in the world, and HBV infection is one of the diseases that seriously affect human health. Chronic HBV infection is closely related to the occurrence of liver fibrosis and liver cancer. However, most of the chronic HBV patients did not get the expected results under conventional treatment. Although interferon can inhibit the replication of HBV, HBV can resist the anti-virus mechanism of the body by inhibiting the secretion and function of interferon, and finally lead the body to a state of immune tolerance. For example, in the peripheral blood and liver infiltrating cells of patients with chronic infection, the proportion of regulatory T cells was significantly increased. In addition, HBsAg, HBeAg, and other viral agents can directly suppress TLR-mediated innate ...

Claims

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Application Information

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IPC IPC(8): C12N15/113A61K48/00A61P1/16A61P31/20
Inventor 田志刚韩秋菊张彩张建
Owner SHANDONG UNIV
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