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Construction and screening as well as applications for siRNAs expression carrier of stomach cancer target STAT3 gene

A technology for expressing vectors and genes, applied in the field of screening, gene therapy drugs, and construction of siRNAs expression vectors, can solve the unsatisfactory survival rate of gastric cancer patients and other problems

Inactive Publication Date: 2012-01-04
济南和合医学检验有限公司
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Problems solved by technology

Despite continuous improvement in surgical treatment and adjuvant chemotherapy, the survival rate of gastric cancer patients is still unsatisfactory

Method used

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  • Construction and screening as well as applications for siRNAs expression carrier of stomach cancer target STAT3 gene
  • Construction and screening as well as applications for siRNAs expression carrier of stomach cancer target STAT3 gene
  • Construction and screening as well as applications for siRNAs expression carrier of stomach cancer target STAT3 gene

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Embodiment Construction

[0082] The present invention will be further described below in conjunction with example, but not limited thereto. All reagents and raw materials of the present invention can be purchased through the market.

[0083] 1. Main Reagents

[0084] 1. The vector pGPU6 / GFP / Neo siRNA Expression Vector was purchased from GenePharma (Catalog no: E-07)

[0085] 2. The engineering bacteria JM109 was purchased from Promega (Catalog no: L1001)

[0086] 3. T4 ligase was purchased from NEW ENGLAND (Catalog no: M0202); plasmid extraction and purification kit (QIAprep Spin Miniprep Kit) was purchased from QIAGEN (Cat No: 27104)

[0087] 4. STAT3 and β-actin-specific PCR primer synthesis, STAT3-specific siRNA oligonucleotide chain synthesis and DNA sequence determination (Nanjing GenScript Biotechnology Co., Ltd.)

[0088] 5. Transfection reagent Lipofectamine 2000 (Cat. No: 11668-027, Invitrogen)

[0089] 6. TRzol Reagent (Cat. No. 152476-026, Invitrogen) RT-PCR Kit RNA PCR Kit (AMV) Ver.3....

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Abstract

The invention relates to construction and screening as well as applications for a siRNA expression carrier of a stomach cancer target STAT3 gene. Two expression plasmids STAT3-siRNA1 / 2 in mammal cells for expressing siRNA are constructed by mainly utilizing an RNA interference technology aiming at different target sequences of an STAT3 gene. The siRNAs expression carrier of the stomach cancer target STAT3 gene provided by the invention can inhibit STAT3 gene expression efficiently with specificity, and can be used for preparing gene medicaments for treating high-expression STAT3 gene stomach cancer.

Description

technical field [0001] The invention relates to gene therapy drugs for gastric cancer expressing STAT3 (signal transducer and activator of transcription 3) gene, in particular to the construction, screening and application of siRNAs expression vectors targeting gastric cancer STAT3 gene, belonging to the technical field of biomedicine. Background technique [0002] Gastric cancer is one of the most common cancers. There are about 930,000 newly diagnosed cases worldwide each year, and about 700,000 patients die from gastric cancer every year. Although surgical treatment and adjuvant chemotherapy techniques have been continuously improved, the survival rate of gastric cancer patients is still unsatisfactory. [0003] In recent years, signal transduction pathways have been paid more and more attention in the study of tumor pathogenesis and prevention methods. Among them, the research on signal transduction pathway of signal transducer and activator of transcription family (STA...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12N15/113A61K48/00A61P35/00
Inventor 肖东杰郏雁飞郑燕童书青马晓丽汪运山
Owner 济南和合医学检验有限公司
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