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Application of an endogenous non-coding small RNA miR-873

A rnamir-873, non-coding technology, applied to the application field of endogenous non-coding small RNA, medicine, can solve the problem of little research on the regulation of target gene mRNA translation process

Active Publication Date: 2018-08-21
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous studies on gene expression and functional changes mainly focused on the transcriptional level and the post-translational modification of gene products, and little research has been done on the regulation of target gene mRNA translation

Method used

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  • Application of an endogenous non-coding small RNA miR-873
  • Application of an endogenous non-coding small RNA miR-873
  • Application of an endogenous non-coding small RNA miR-873

Examples

Experimental program
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Effect test

Embodiment 1

[0026] (1) To study the dynamic changes of miR-873 in the spinal cord dorsal horn and dorsal root ganglion (DRG) during the formation of morphine tolerance

[0027] Exiqon's LNA (locked nucleic acid) technology microRNA expression profile chip was used to detect chronic morphine tolerance group (NS group) and control group ( Sham group) miRNAs differentially expressed in the lumbar spinal cord dorsal horn and DRG. The pre-experiment has been completed and the microarray detection of differential expression profiles in the dorsal horn of the lumbar spinal cord on the 1st day after the formation of morphine tolerance has been completed. Using stem-loop reverse transcription primer real-time fluorescent quantitative PCR (stem-loop RT-qPCR, ABI company, TaqMan probe method) to verify the expression of miRNAs, first use miR sequence-specific stem-loop reverse transcription primer for reverse transcription, Then qPCR was performed in combination with miR-specific sense and antisens...

Embodiment 2

[0039] The specific process of lentivirus-mediated miR-873 interference vector construction:

[0040] According to the miR-873 sequence, using the shRNA design program, according to the principle of RNA interference sequence design, design the interference target sequence 5'-AUAAGGAUUUUUAGGGCAUUA-3'; use EcoRI and BamHI as enzymes at both ends of the pLenR-GPH vector (Jikai Gene) Carry out enzyme digestion at the cleavage site to make it linear, perform a ligation reaction with the double-stranded interference target sequence by T4DNA ligase to form an shRNA expression vector, and transfer the ligated product into the prepared competent Escherichia coli. The clones were identified by enzyme digestion, the positive clones were selected for sequencing, and after sequencing and comparison, the positive clones were identified as the successfully constructed shmiR-873 interference vectors;

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Abstract

The invention provides an application of endogenic non-coding small RNA miR-873, particularly an application of the miR-873 in drugs for treating or alleviating chronic pain and morphine tolerance. In addition, the endogenic non-coding small RNA miR-873 also can be used in the preparation of preparations for regulating the A20 gene and regulating preparations for the proinflammatory factor TNF Alpha released by the spinal cord under the condition of morphine tolerance. The invention proves that since the miR-873 can regulate the TNF alpha-induced protein-3 (A20) to participate in the development of morphine tolerance, an important foundation is laid for the application of the miR-873 in drugs for treating or alleviating chronic pain and morphine tolerance.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. The present invention relates to the use of an endogenous non-coding small RNA, in particular to the use of miR-873 in the preparation of medicines for treating or alleviating chronic pain and morphine tolerance. Background technique [0002] Morphine, as a classic opioid analgesic drug, is widely used in the clinical treatment of acute and chronic pain. However, morphine tolerance (Morphine tolerance, MT) caused by long-term use of morphine is a major medical problem that limits its clinical application. Because the mechanism of morphine resistance is not fully understood, and there is a lack of effective prevention and treatment measures, morphine resistance has always been a hot spot in the field of chronic pain research. [0003] MicroRNA (miRNA, miR) is a kind of non-coding single-stranded small molecule RNA with a length of about 21-23 nucleotides, which consists of a single-stranded ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61P29/00
Inventor 邹望远王健郭曲练宋宗斌
Owner CENT SOUTH UNIV
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