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Human miR-210 biological knock-down element, construction method and application thereof

A technology of mir-210 and construction method, which is applied in the field of human miR-210 biological knockdown elements to facilitate scientific research and control the malignant phenotype and biological behavior of tumors

Inactive Publication Date: 2016-08-24
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Abstract
  • Description
  • Claims
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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-210 biological knock-down element, construction method and application thereof
  • Human miR-210 biological knock-down element, construction method and application thereof
  • Human miR-210 biological knock-down element, construction method and application thereof

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Embodiment Construction

[0025] The present invention will be further described in detail below through specific embodiments in conjunction with the accompanying drawings.

[0026] Technology and method:

[0027] 1. Cell culture

[0028] 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.

[0029] 2. Construction of miR-210 biological knockdown element

[0030] The designed antisense sequence of human miR-210 was chemically synthesized in vitro; and a connecting sequence was added between the two antisense sequences to finally constitute 6 copies of the knockdown element. 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. The two were ligated by T4 DNA liga...

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Abstract

The invention discloses a human miR-210 biological knock-down element, a construction method and an application thereof. Sequence of the biological knock-down element is as shown in the SEQ ID NO:1. Abnormal expression of miR-210 exists in many tumor cells, and the family participates in regulation of cell proliferation, apoptosis and cell migration related genes. Through design of the knock-down element, expression of miR-210 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 biological knockdown element of human miR-210, its construction method and application. Background technique [0002] As an important representative of non-coding RNA, microRNA (miRNA) has attracted more and more attention from researchers. miRNA is a class of non-coding RNA molecules with a length of about 19-23nt. They generally inhibit protein translation or cause mRNA degradation by binding to the 3'-UTR of mRNA. miRNA plays a huge role in the regulation of phenotypes such as cell proliferation, apoptosis, and differentiation, as well as in the development of organisms and the occurrence and development of diseases. Therefore, the understanding of miRNA function is very important for the research of cell biology and the diagnosis and treatment of diseases. [0003] miR-210 is the most significantly overexpressed miRNA in hypoxic tumor cells, and plays an important role in the proliferation, apoptosis, angiogen...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/10A61K48/00A61P35/00A61P13/10C12Q1/68
Inventor 刘宇辰黄卫人韩永华蔡志明
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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