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Small antisense oligonucleotide aiming at miR-92a seed sequence and application

A technology of mir-92a and oligonucleotides, which is applied in tiny antisense oligonucleotides and its application fields, can solve problems such as interference and pairing, and achieve the effects of strong specificity, small molecular weight, and high transfection efficiency

Inactive Publication Date: 2015-04-22
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Antisense nucleic acids targeting miRNA seed sequences can interfere with the pairing of miRNAs with target genes, thereby preventing miRNAs from functioning

Method used

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  • Small antisense oligonucleotide aiming at miR-92a seed sequence and application
  • Small antisense oligonucleotide aiming at miR-92a seed sequence and application
  • Small antisense oligonucleotide aiming at miR-92a seed sequence and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Design and synthesis of tiny antisense oligonucleotides

[0023] The seed sequence of human microRNA-92a was obtained from the microRNA Families database, and a small antisense oligonucleotide sequence (t-antimiR-92a) targeting the seed sequence of human microRNA-92a was designed according to the principle of sequence complementarity, and analyzed and determined by BLAST software Its small antisense oligonucleotide sequence and random control sequence ( figure 1 ). Tiny antisense oligonucleotide sequence: t-antimiR-92a (5'-AGTGCAAT-3'), random sequence (scramble, SCR): 5'-TCATACTA-3'; all synthesized by Shanghai Bioengineering Co., Ltd., All sulfur modification, high performance liquid chromatography (high performance liquid chromatography, HPLC) purification.

Embodiment 2

[0024] Example 2 Cell Culture

[0025] RPMI-8266 cells (multiple myeloma cells, purchased from the Shanghai Institute of Cell Biochemistry, Chinese Academy of Sciences) were inoculated in RPMI-1640 medium containing 10% fetal bovine serum by volume fraction and antibiotic-free, and placed at 37°C, volume The fraction is 5% CO2 incubator, cultivated under saturated humidity. Change the medium every 2 to 3 days for passage. Cells in the logarithmic growth phase with a mass fraction of 0.2% trypan blue and an exclusion rate of >95% were selected for the experiment.

Embodiment 3

[0026] Example 3 Signal pathway analysis of miRNA-92a

[0027] miRFocus (http: / / mirfocus.org) software can be used to analyze the target genes of miRNA and the signaling pathways involved.

[0028] miRFocus software analyzed the signaling pathways involved in miR-92a. The result is as figure 2 , miR-92a participates in many important signaling pathways related to tumors. Such as mTOR signaling pathway, cell cycle and other cancer pathways, etc.

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Abstract

The invention belongs to the field of medicine preparation, and particularly relates to a small antisense oligonucleotide aiming at a miR-92a seed sequence and an application. A nucleotide sequence of the small antisense oligonucleotide is 5'-AGTGCAAT-3'; the small antisense oligonucleotide is subjected to perthio modification, targets on miR-92a in multiple myeloma cells RPMI-8266, and has an inhibiting effect on the multiple myeloma cells, so that the small antisense oligonucleotide can be applied to preparation of medicines for preventing or treating tumors, and has a wide prospect in clinical treatment.

Description

technical field [0001] The invention belongs to the field of medicine preparation, and in particular relates to a small antisense oligonucleotide for miR-92a seed sequence and its application. Background technique [0002] Multiple myeloma (multiple myeloma, MM) is a monoclonal malignant plasma cell disease. Thalidomide, lenalidomide, and bortezomib, which appeared recently, can effectively prolong the life of MM patients, but drug resistance or disease recurrence is still a concern. The pathogenesis of multiple myeloma involves abnormal expression of multiple genes and dysregulation of multiple signaling pathways. Studies have found that these dysregulated genes and signaling pathways can be regulated by miRNAs. [0003] microRNA (miRNA) is a kind of non-coding RNA, which can complementarily bind to the 3' non-coding region of the target mRNA to inhibit the translation of mRNA or degrade the mRNA chain. Therefore, MiRNA can affect various functions of cells by affecting ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K48/00A61P35/00
Inventor 费嘉丰茂晓骆小闯古春明阴钊
Owner JINAN UNIVERSITY
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