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ShRNA for inhibiting human EDARADD gene expression, lentiviral vector and construction method and application of lentiviral vector

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

Active Publication Date: 2020-02-04
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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  • ShRNA for inhibiting human EDARADD gene expression, lentiviral vector and construction method and application of lentiviral vector
  • ShRNA for inhibiting human EDARADD gene expression, lentiviral vector and construction method and application of lentiviral vector
  • ShRNA for inhibiting human EDARADD gene expression, lentiviral vector and construction method and application of lentiviral vector

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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 shRNA for inhibiting human EDARADD gene expression, a lentiviral vector and a construction method and application of the lentiviral vector. The method comprises the following steps: firstly, synthesizing a positive strand and a negative strand of DNA oligo aiming at an RNA interference target sequence; annealing the positive strand and the negative strand to form double-stranded DNA with a cohesive end; linearizing the lentiviral vector through enzyme digestion, and connecting the linearized vector with double-stranded DNA; transforming escherichia coli competent cellswith a connection product, and carrying out PCR identification on the obtained positive clone; and performing plasmids extraction for virus packaging in the next step. According to the invention, theRNA interference target sequence is designed by taking an EDARADD gene as a template, an shRNA interference sequence is designed according to a selected target sequence, an RNA interference lentiviralvector is constructed through packaging, and after the EDARADD gene is knocked down, proliferation of tongue squamous cell carcinoma cells CAL-27 can be remarkably influenced, the cell clone formingcapacity is influenced, and cell apoptosis is promoted.

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 ...

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Application Information

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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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