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Construction method and pharmaceutical use of microRNA adenovirus expression plasmid for severe hepatitis related gene hfg12, hFas and hTNFR1

A technology for expressing plasmids and severe hepatitis, applied in the direction of double-stranded DNA virus, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of lack of specific and effective treatment methods, dangerous disease, liver cell death, etc.

Active Publication Date: 2009-01-21
武汉大风生物科技有限公司
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Problems solved by technology

[0006] Severe hepatitis can be caused by a variety of etiologies clinically, and its onset is dangerous. Patients may experience large areas of liver cell death in a short period of time. Currently, there is no specific and effective treatment

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  • Construction method and pharmaceutical use of microRNA adenovirus expression plasmid for severe hepatitis related gene hfg12, hFas and hTNFR1
  • Construction method and pharmaceutical use of microRNA adenovirus expression plasmid for severe hepatitis related gene hfg12, hFas and hTNFR1
  • Construction method and pharmaceutical use of microRNA adenovirus expression plasmid for severe hepatitis related gene hfg12, hFas and hTNFR1

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Embodiment Construction

[0067] The present invention will be further described in detail below in conjunction with specific embodiments.

[0068] The present invention constructs the miRNA expression plasmid of hfgl2 prothrombinase, hFas and hTNFR1, and confirms its interference effect at the cell level, including the following steps (taking the construction of hFas-microRNA expression plasmid as an example):

[0069] 1. Construction of pcDNA expression vector

[0070] Construction of hFas-miRNA expression plasmid: According to the hFas cDNA sequence on PubMed, use the miRNA design software of Invitrogen Company to design and synthesize 3 pairs of complementary oligonucleotide single strands. According to the molecular cloning procedure, insert ds Oligo into the microRNA expression vector pcDNA TM 6.2-GW / EmGFP-miR, construct three hFas-miRNA plasmids: p-hFasmi-RNA1, p-hFasmiRNA2 and p-hFasmiRNA3. The miRNA expression plasmids of the unmutated hFas gene were screened by sequencing.

[0071] 2. Expe...

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Abstract

The invention relates to a group of genetic biological agent, namely microRNA adenovirus expression plasmids which construct severe hepatitis related human fgl2 (Human Fibrinogen-like protein 2, hfgl2) prothrombinase, Fas and tumor necrosis factor recipient I (tumor necrosis factor, TNFR1) genes. The effectiveness and the specificity of microRNA interference plasmids are detected in the cellular level and three kinds of the microRNA adenovirus expression plasmids are constructed and applied together to treat severe hepatitis. The group of the genetic biological agent adopts pAd / CMV / V5-DEST vectors and pcDNA expression plasmids of the hfgl2, the hFas and the hTNFR1 to construct pAd-hfgl2, pAd-hFas and pAd-hTNFR1 through the Gateway technology.

Description

technical field [0001] The present invention relates to the construction of a group of gene biological preparations, that is, the construction of human fgl2 (Human Fibrinogen-like protein 2, hfgl2) prothrombinase, Fas and tumor necrosis factor receptor I (tumor necrosis factor, TNFR1) genes related to severe hepatitis The microRNA adenovirus expression plasmid is combined with three plasmids for treatment of severe hepatitis, and its pharmaceutical use is verified. Background technique [0002] Due to the high incidence and great harm of viral hepatitis, it has become an important disease that seriously affects people's health and living standards and restricts economic development. So far, there is no confirmed effective treatment for severe hepatitis in the world. Exploring the pathogenesis and prevention methods of severe hepatitis is an important topic in the field of applied basic and clinical research. [0003] Infecting different strains of inbred mice with mouse hep...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/861C12N15/11C12N15/70A61K48/00A61P1/16A61P31/12C12N15/113
CPCC12N15/86C12N15/1138C12N15/1137C12N2330/51C12N2710/10343C12N7/00C12N2310/141C12N2799/02A61P1/16A61P31/12
Inventor 宁琴罗小平习东高随朱传龙郭健文严伟明王鸣
Owner 武汉大风生物科技有限公司
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