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Construction and screening of siRNAs (small interfering RANs) expression vectors of gastric cancer targeted DEC1 (differentiated embryo-chondrocyte expressed gene 1) and application thereof

A technology of DEC1 and expression vectors, which is applied in the fields of screening, gene therapy drugs, and construction of siRNAs expression vectors, and can solve problems such as procaspase8 not working

Inactive Publication Date: 2012-07-18
济南赛尔生物科技股份有限公司
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Problems solved by technology

DEC1 can resist serum starvation-induced apoptosis and selectively inhibit the activity of procaspases 3, 7, 9 in colon cancer cells, but has no effect on procaspase 8

Method used

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  • Construction and screening of siRNAs (small interfering RANs) expression vectors of gastric cancer targeted DEC1 (differentiated embryo-chondrocyte expressed gene 1) and application thereof
  • Construction and screening of siRNAs (small interfering RANs) expression vectors of gastric cancer targeted DEC1 (differentiated embryo-chondrocyte expressed gene 1) and application thereof
  • Construction and screening of siRNAs (small interfering RANs) expression vectors of gastric cancer targeted DEC1 (differentiated embryo-chondrocyte expressed gene 1) and application thereof

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Embodiment Construction

[0078] The present invention will be further described below in conjunction with embodiment, but not limited thereto. All reagents and raw materials of the present invention can be purchased through the market.

[0079] 1. Main Reagents

[0080] 1. The vector pGreenPuro shRNA Cloning and Expression vector was purchased from SBI (Catalog no: SI505A-1)

[0081] 2. The engineering bacteria JM109 was purchased from Promega (Catalog no: L1001)

[0082] 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)

[0083] 4. Synthesis of DEC1 and β-actin-specific PCR primers, primers ForwardH PCR Primer and ReverseH PCR Primer, DEC1-specific siRNA oligonucleotide chain synthesis and DNA sequence determination (Nanjing KingScript Biotechnology Co., Ltd.)

[0084] 5. Transfection reagent Lipofectamine 2000 (Cat. No: 11668-027, Invitrogen)

[0085] 6. TRzol Reagent (Cat...

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Abstract

The invention relates to construction and screening of siRNAs (small interfering RANs) expression vectors of a targeted DEC1 (differentiated embryo-chondrocyte expressed gene 1) resulting in gastric cancer and application thereof. In the invention, by mainly adopting an RNA interference technology, two expression plasmids DEC1-siRNA1 / 2 capable of expressing siRNA in mammalian cells are constructed by aiming at different target sequences of the DEC1. The siRNAs expression vectors of the targeted DEC1 resulting in gastric cancer can efficiently and specifically inhibit DEC1 expression; and the siRNAs expression vectors are used for preparing a gene drug for treating gastric cancer resulting from the high expressed DEC1.

Description

technical field [0001] The invention relates to gene therapy drugs for gastric cancer expressing DEC1 (Differentiated embryo-chondrocyte expressed gene 1) gene, in particular to the construction, screening and use of siRNAs expression vectors targeting gastric cancer DEC1 gene, belonging to the technical field of biomedicine. Background technique [0002] Differentiated embryo-chondrocyte expressed gene 1 (DEC1) is a new member of basic helix-loop-helix (bHLH) protein family, which contains bHLH domain. Recently, it has been found that it is a transcription factor closely related to the occurrence and development of tumors. Its function involves the proliferation, differentiation, apoptosis and premature aging of tumor cells. It is a gene involved in the regulation of hypoxia and is closely related to the hypoxic state of tumor tissues. It can be used as a predictive marker of tumor response after radiotherapy and chemotherapy. Studies have found that DEC1 can resist serum ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/85C12N15/113A61K48/00A61P1/00A61P35/00
Inventor 王恒桓郏雁飞郑燕马晓丽肖东杰孙善会魏炎汪运山蔡平平
Owner 济南赛尔生物科技股份有限公司
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