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Single-stranded specific shRNA for human fibrinogen, recombinant interference vector and application

A human fibrinogen and fibrin technology, applied in the direction of reverse transcription RNA virus, DNA/RNA fragment, application, etc., can solve the problems of biological function tumor type difference, imbalance, reduce specific fibrin and other problems, and achieve enhanced treatment effect of sensitivity

Pending Publication Date: 2020-09-11
刘玉琳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the unbalanced expression of FGA, FGB and FGG levels in different tumor cells, there are also differences in tumor types in their biological functions.
Whether siRNA can be used to reduce the level of specific fibrinogen single chain to achieve the purpose of improving the therapeutic effect of tumors has not yet been reported

Method used

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  • Single-stranded specific shRNA for human fibrinogen, recombinant interference vector and application
  • Single-stranded specific shRNA for human fibrinogen, recombinant interference vector and application
  • Single-stranded specific shRNA for human fibrinogen, recombinant interference vector and application

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Embodiment Construction

[0038] The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0039] Specific experimental methods not specified in the examples were performed according to conventional experimental conditions, referring to the experimental protocol described in the "Molecular Cloning Experiment Guide (Third Edition, J. Sambrook et al.)" or the conditions suggested by the manufacturer.

[0040] 1. Acquisition of specific interference sequences for each single chain (FGA, FGB and FGG) of human fibrinogen:

[0041] siRNA sequences were designed according to the single-strand sequences of human fibrinogen published by NCBI (FGA, NM_000508.5; FGB, NM_005141.5; FGG, NM_000509.5), and the specific design method was to utilize the RNAi online design tool of the Life Technologies website ( https: / / rnaidesigner.thermofisher.com / rnaiexpress / sort.do ) Design the interference target sequence according to the human fibrinogen si...

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Abstract

The invention relates to single-stranded specific shRNA for human fibrinogen, a recombinant interference vector and an application. For single strands of the human fibrinogen, specific interference RNA and the recombinant interference vector expressing shRNA are designed, tumor cells are subjected to transfection, the expression of fibrinogen of tumor cells is knocked down, and sensibility of tumors to chemotherapy medicines and targeting medicines is reinforced. The human fibrinogen shRNA disclosed by the invention can specifically reduce the level of the fibrinogen, and through constructingthe shRNA recombinant interference vector and performing slow virus packaging, the expression of the fibrinogen interfering the tumor cells can be stabilized. Experiments prove that through knocking down single strands of the fibrinogen, the treatment sensibility of different types of tumors (kidney cancer, ovarian cancer, breast cancer, liver cancer, colon cancer, nasopharyngeal carcinoma, gastric cancer or lung cancer) can be reinforced, and the single-stranded specific shRNA is an important target point for reinforcing traditional treatment sensibility of the tumor.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a specific shRNA of human fibrinogen single chain, a recombinant interference carrier and applications. Background technique [0002] Human fibrinogen, also known as human coagulation factor I (Human coagulation factor I), is a polymeric peptide composed of two protein trimers, which is composed of two groups of Aα, Bβ and γ protein peptide chains connected by 29 disulfide bonds. The three single-chain proteins are respectively transcribed by independent mRNAs. The related genes are all located on the long arm of chromosome 4, q23-q32, with a total length of 50kb. They are a group of discontinuous single-copy genes. Fibrinogen is a macromolecular glycoprotein that is released around cells through cellular secretion. Cancer patients, especially those with advanced cancer, often present hyperfibrinogenemia. The applicant's previous basic research found that the ectopic expression of tu...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/867C12N15/66A61K31/713A61P35/00
CPCC12N15/113C12N15/86C12N15/66A61K31/713A61P35/00C12N2310/14C12N2320/30C12N2740/15043
Inventor 刘玉琳王强颜泽萱徐丽丽
Owner 刘玉琳