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Construction method and application of human miR-106b-5p interference fragment

A technology of mir-106b-5p and hsa-mir-106b-5p, which is applied in the field of biotechnology and genetic engineering, can solve the problems that the role of miR-106b-5p is not clear and not fully elucidated, and achieve the inhibition of human skin melanoma The effect of cell proliferation

Pending Publication Date: 2019-05-21
信雅生物科技(苏州)有限公司
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Problems solved by technology

However, the role of each member of the miR-106b-25 cluster in tumors has not yet been fully elucidated.
More importantly, the role of miR-106b-5p in the malignant progression of melanoma remains unclear

Method used

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  • Construction method and application of human miR-106b-5p interference fragment
  • Construction method and application of human miR-106b-5p interference fragment
  • Construction method and application of human miR-106b-5p interference fragment

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Embodiment Construction

[0054] A method for constructing a miR-106b-5p interference nucleotide sequence that effectively inhibits melanoma cell proliferation, comprising the steps of:

[0055] (1) Using the miRbase (http: / / www.mirbase.org / ) database, obtain the nucleotide sequence of mature hsa-miR-106b-5p (hsa-miR-106b-5p MIMAT0000680), the specific sequence is as follows:

[0056] UAAAGUGCUGACAGUGCAGAU

[0057] (2) Use Primer Premier 5.0 software to design primers. In addition to meeting the primer design principles, the designed primers should include the full-length hsa-miR-106b-5p interference sequence in the PCR product. The specific sequence of hsa-miR-106b-5p interference sequence is as follows:

[0058] CGUUCAUGGUGUCACGC

[0059] (3) Using a chemical synthesis method to synthesize RNA single strands.

[0060] (4) In order to verify the application of hsa-miR-106b-5p interference fragment in skin melanoma cells, we detected the expression of hsa-miR-106b-5p in melanoma tissue and its benig...

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Abstract

Since the death rate of melanoma is high, by developing a novel molecular therapeutic target for inhibiting cell proliferation of the melanoma, a laboratory basis for clinical treatment of the melanoma can be provided. MicroRNA (micro-RNA, for short, MiRNA) is non-coding micromolecular RNA of which the length is about 22 nucleotides, plays an important role in the cell proliferation process, has the advantages of being high in tissue specificity, not easy to degrade, easy to detect and the like, and is an ideal target of molecular targeted therapy. Therefore, the invention provides a construction method of a human miR-106b-5p (hsa-miR-106b-5p) interference fragment and application of the human miR-106b-5p interference fragment in the cell proliferation of the melanoma, which can provide anew experimental evidence for basic research and clinical treatment of the melanoma. The full name of miR-106b-5p is human micro ribonucleic acid-106b-5`end (hsa-miR-106b-5p), and the former name of the miR-106b-5p is hsa-miR-106b.

Description

technical field [0001] The invention belongs to the field of biotechnology and genetic engineering, and in particular relates to a construction method and application of an interference fragment of human tiny miR-106b-5p (hsa-miR-106b-5p) that inhibits the proliferation of melanoma cells. Background technique [0002] MicroRNA (miRNA) is a kind of non-coding small molecule RNA with a length of about 22 nucleotides, which is processed by Dicer enzyme from a single-stranded RNA precursor (pre-miRNA) with a hairpin structure of about 70-90 bases generated after. It degrades mRNA or inhibits its translation by completely or incompletely complementary pairing with the 3' non-coding region (3'UTR) of the target mRNA molecule, and plays an important role in life activities such as individual development, cell apoptosis, proliferation and differentiation. miRNA can regulate the expression of multiple proteins at the post-transcriptional level, and a single target protein can also b...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/10A61K31/7088A61P35/00
Inventor 宋鑫李真孟旭东宋登燕
Owner 信雅生物科技(苏州)有限公司
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