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Human miR-183 gene cluster biological knock-down element, construction method and application thereof

A mir-183 and construction method technology, applied in the field of human miR-183 gene cluster biological knockdown elements, to achieve the effect of controlling tumor malignant phenotype and biological behavior and facilitating scientific research

Inactive Publication Date: 2016-08-24
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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Problems solved by technology

[0004] Although the traditional view is that miRNAs can down-regulate the expression of target genes, recent studies suggest that target genes can also in turn inhibit the expression / activity of miRNAs

Method used

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  • Human miR-183 gene cluster biological knock-down element, construction method and application thereof
  • Human miR-183 gene cluster biological knock-down element, construction method and application thereof
  • Human miR-183 gene cluster biological knock-down element, construction method and application thereof

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Embodiment Construction

[0024] The present invention will be further described in detail below through specific embodiments in conjunction with the accompanying drawings.

[0025] Technology and method:

[0026] 1. Cell culture

[0027] Bladder cancer cell lines T24 and UM-UC-3 were purchased from American Type Culture Collection (ATCC, Manassas, USA), and the cell culture medium consisted of 90% DMEM (Invitrogen, CA), 10% fetal bovine serum (Invitrogen), 1 %-2% glutamine and 0.5%-1% double antibody.

[0028] 2. Construction of biological knockdown elements of miR-183 / 96 / 182 gene cluster

[0029] Two antisense sequences of designed human miR-183, 96, and 182 were chemically synthesized in vitro; and linking sequences were added between these antisense sequences to finally constitute six copies of knockdown elements. Introduce restriction sites Xhol and Notl upstream and downstream of the element, and simultaneously use Xhol and Notl to digest the psiCHECKTM-2 dual luciferase expression vector. Th...

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Abstract

The invention discloses a human miR-183 gene cluster biological knock-down element, a construction method and an application thereof. Sequence is as shown in the SEQ ID NO:1. Abnormal expression of miR-183 gene cluster exists in many tumor cells, and the family participates in regulation of cell proliferation, apoptosis and cell migration related genes. Through design of the gene cluster knock-down element, expression of a gene cluster in bladder cancer cell can be effectively regulated. The knock-down element of the invention is convenient for scientific research and is also helpful for controlling tumor malignant phenotype and biological behavior.

Description

technical field [0001] The present application relates to a human miR-183 gene cluster biological knockdown element, its construction method and application. Background technique [0002] MicroRNAs (miRNAs) are endogenous non-coding RNAs, approximately 22 nucleotides in length. miRNA usually binds to the 3' untranslated region (3'untranslated region, 3'UTR) of its target gene mRNA and negatively regulates its expression. miRNAs can regulate cell proliferation, differentiation and apoptosis. Therefore, the discovery of miRNA provides a new idea for the study of tumor pathogenesis, and provides a new strategy for tumor diagnosis and treatment. [0003] The miR-183 gene cluster (miR-183 family) is a highly conserved gene cluster, including miR-183, miR-182 and miR-96 located on chromosome 7. Studies have shown that there is abnormal expression of miR-183 gene cluster in a variety of tumor cells, and this family is involved in the regulation of cell proliferation, apoptosis, ...

Claims

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Application Information

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IPC IPC(8): C12N15/113A61K48/00A61P35/00C12Q1/68
Inventor 黄卫人刘宇辰韩永华蔡志明
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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