Targetted liver cancer AFP gene siRNAs expression carrier and its constructing method and use

A technology for expressing vectors and genes, used in the fields of siRNAs expression plasmids and gene therapy drugs

Inactive Publication Date: 2006-04-05
JINAN CENTER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

In addition, antisense nucleic acid molecules can only achieve the effect of inhibiting gene expression at a relatively high concentration, and its price and mass production are also a restrictive factor

Method used

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  • Targetted liver cancer AFP gene siRNAs expression carrier and its constructing method and use
  • Targetted liver cancer AFP gene siRNAs expression carrier and its constructing method and use
  • Targetted liver cancer AFP gene siRNAs expression carrier and its constructing method and use

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Embodiment Construction

[0071] The present invention will be further described below in conjunction with embodiment, but not limited thereto.

[0072] 1. Materials used for the preparation of siRNAs expression vector targeting liver cancer AFP gene

[0073] 1.1 Plasmids and strains Plasmid pSilencer 3.0-H1 was purchased from Ambion, see figure 1 , the engineering strain DH5α was purchased from the Institute of Molecular Biology of Shandong University.

[0074] 1.2 Enzymes and main reagents T4 DNA ligase was purchased from Promega Company in the United States. The extraction and purification of plasmids were prepared by classic alkaline lysis method and polyethylene glycol plasmid purification method.

[0075] 1.3 Sequencing primers M13 universal primers were used for sequencing the inserts in the plasmids. Forward Primer: 5'-CACGACGTTGTAAAAGAC-3', Reverse Primer: 3'-GGACACATTTAACAATAG G-5', purchased from Shanghai Shenyou Company.

[0076] 1.4 Cell line and medium: The liver cancer cell line SMMC-...

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Abstract

This invention, in biopharmaceutical technology, relates to target liver cancer AFP genetic siRNAs expression vehicle and its establishment and use. It is used to prepare genetic medicines secreting AFP protein. With pSilencer3.0-H1 as carrier, expression of hairpin double chain RNA molecule is ignited by H1 against AFP gene, with plasmid of the said vehicle against AACTCAGTGAGGACAAACTAT-3' of AFP gene encoding region sequence 5', which is material used for treating liver cancer.

Description

(1) Technical field [0001] The invention relates to a gene therapy drug for liver cancer expressing AFP gene, in particular to an siRNAs expression plasmid targeting the AFP gene of liver cancer, and belongs to the technical field of biomedicine. (2) Background technology [0002] Primary liver cancer (Primary hepatocellular carcinoma, PHCC) is one of the most common malignant tumors, and its average survival period is only about 4 months from the establishment of diagnosis. It is a disease that seriously endangers human health. Although many methods have been used to treat primary liver cancer, such as chemotherapy, radiotherapy, surgery, biological therapy, etc., none of them can significantly improve the survival rate of patients with primary liver cancer. Therefore, finding effective treatment means has very important social and economic benefits. [0003] Alpha fetoprotein (AFP) is the main serum protein produced by the liver and yolk sac during the embryonic period of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12N15/85C12N15/12C12N15/10C12Q1/68A61K48/00A61P35/00
Inventor 汪运山亓同钢
Owner JINAN CENTER HOSPITAL
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