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A kind of shrna that suppresses human edaradd gene expression, lentiviral vector and its construction method and application

A technology of lentiviral vector and gene expression, applied in the direction of retroRNA virus, DNA/RNA fragment, virus/bacteriophage, etc.

Active Publication Date: 2021-02-19
XUZHOU CENT HOSPITAL
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Problems solved by technology

However, the molecular targeted therapy of tongue squamous cell carcinoma is still in the experimental research stage, and the correct selection of specific molecular targets is very important
[0004] Whether the human EDARADD gene is related to tumor cell apoptosis and its role in tumorigenesis and progression has not been reported in the literature

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  • A kind of shrna that suppresses human edaradd gene expression, lentiviral vector and its construction method and application
  • A kind of shrna that suppresses human edaradd gene expression, lentiviral vector and its construction method and application
  • A kind of shrna that suppresses human edaradd gene expression, lentiviral vector and its construction method and application

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Embodiment Construction

[0036] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0037] The main indicators for evaluating the biological behavior of tumors are tumor cell proliferation, clone formation ability, apoptosis and so on. In this study, by knocking down the expression of endogenous EDARADD gene in tongue squamous cell carcinoma cell line, we observed its effect on the biological behavior of tumor cells, so as to provide reference for prognosis judgment, surgery and targeted therapy of tongue squamous cell carcinoma patients. Exploring a non-invasive, safe and convenient way to detect the specific gene EDARADD will help in the diagnosis, treatment and prognosis prediction of tongue squamous cell carcinoma.

[0038] 1. Gene information

[0039]The online resource UALCAN database (http: / / ualcan.path.uab.edu / ) generated boxplots of the expression levels of EDARADD in head and neck tumor tissues as well as ...

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Abstract

The invention discloses a shRNA, a lentiviral vector and its construction method and application for inhibiting the expression of human EDARADD gene. Firstly, the forward strand and the reverse strand of the DNA oligo aiming at the RNA interference target sequence are synthesized; the forward strand and the reverse strand are annealed to form a band Double-stranded DNA at the cohesive end; linearize the lentiviral vector by enzyme digestion, connect the linearized vector to the double-stranded DNA; transform the ligated product into E. coli competent cells, and perform PCR identification on the positive clones obtained; plasmid extraction, For the next step of virus packaging. The present invention uses the EDARADD gene as a template to design the RNA interference target sequence, designs the shRNA interference sequence according to the selected target sequence, packages and constructs the RNA interference lentiviral vector, and knocks down the EDARADD gene, which will significantly affect the CAL of tongue squamous cell carcinoma cells Proliferation of ‑27 cells, affecting cell clone formation ability, and promoting cell apoptosis.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and biomedicine, and in particular relates to a shRNA and a lentiviral vector for inhibiting the expression of human EDARADD gene and its construction method and application. Background technique [0002] Tongue squamous cell carcinoma (TSCC), referred to as tongue squamous cell carcinoma, is one of the most common malignant tumors in the oral and maxillofacial region, with high mortality and morbidity. Its development is a multistep process involving genetic alterations and epigenetic modifications. TSCC is a causative agent of many variables, accumulation of multiple genetic alterations and environmental factors, such as tobacco use, alcohol consumption, chronic inflammation, and human papillomavirus (HPV) infection, among others, and several recent reports suggest that TSCC in young adults increases in incidence , especially in women, the rate of distant metastasis is higher and the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/867C12N15/66A61K31/713A61P35/00
CPCA61P35/00C12N15/1138C12N15/66C12N15/86C12N2310/14C12N2740/15043C12N2800/107C12N2310/531
Inventor 孟箭李梦白玉婷李晓东管俊杰王守鹏
Owner XUZHOU CENT HOSPITAL
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