Circular RNA circ-01477 and application thereof

A technology of circ-01477 and rnacirc-01477, applied in the field of nerve regeneration and repair

Active Publication Date: 2020-02-21
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the role of circRNAs in the repair of central nervous system injury has been rarely reported so far, especially the phenotype regulation of astrocytes

Method used

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  • Circular RNA circ-01477 and application thereof
  • Circular RNA circ-01477 and application thereof
  • Circular RNA circ-01477 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Expression and identification of circ-01477

[0024] RNA-seq was performed on the spinal cord tissues of SD (Sprague Dawley) rats at different time points (0d, 1d, 3d, 7d, 14d, 21d and 28d) after spinal cord hemisection injury. According to the sequencing results, select circRNAs that are all composed of exons and have significantly different expressions of loops >500nt. The sequencing results analyzed the screened circRNAs, and further verified the existence of circRNAs by PCR. According to the design principles of circRNAs primers, back primers were designed for these circRNAs, and NCBI was used to design primers for each circRNA. Select different annealing temperatures according to the primer information for PCR amplification, and then select circRNAs with the expected band size through agarose gel electrophoresis, send them to the company for sequencing, and compare the sequencing results with the original sequence to verify whether they span the circula...

Embodiment 2

[0048] Example 2: Functional verification of circRNA

[0049] 1. Culture of primary astrocytes

[0050] The spinal cords of red-skinned mice were isolated one day after birth, and transferred to a petri dish filled with isolation buffer (DMEM). The meninges were peeled off under a stereoscope to separate the spinal cord, and transferred to another Petri dish filled with separation buffer (DMEM). The tissue was washed with separation buffer (DMEM / F12), and the tissue was minced with surgical scissors. Add 2ml of 0.25% trypsin in the digestive solution, digest at 37°C for 15min, and shake several times every 5min. Use digestion termination solution (DMEM / F12+10% FBS) to terminate the digestion, centrifuge at 1000rpm for 5min, and discard the supernatant. Add complete medium (DMEM / F12+10% FBS+1% p.s+1% L-glu) to resuspend the cells. Microscopically observe the agglomeration of nerve cells, pass through a cell sieve, and collect the cell suspension. After counting, they were pl...

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Abstract

The invention discloses circular RNA circ-01477 and an application thereof to preparation of drugs for promoting nerve regeneration and repairing nerve injuries. Research results indicate that by means of down-regulation or suppression of organism circ-01477 expression, proliferation and migration of astrocytes are suppressed, and repair of central nerve injuries is facilitated.

Description

technical field [0001] The invention belongs to the technical field of nerve regeneration and repair, and specifically relates to the application of circular RNA circ-01477 in the preparation of drugs for promoting nerve regeneration and repairing nerve damage. Background technique [0002] Spinal cord injury is a serious type of nerve injury, which can lead to permanent movement and sensory disturbances and affect people's quality of life. After spinal cord injury, the body will undergo a series of changes at the cellular and molecular levels, involving a variety of cells, including astrocytes, oligodendrocytes, microglia, vascular endothelial cells, etc. environment. Among them, astrocytes proliferate after spinal cord injury, and then form glial scars at the injury site, hindering axon regeneration and neural circuit remodeling. In addition, glial scars can also limit the spread of inflammatory responses to surrounding tissues, repair the damaged blood-brain barrier, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K31/713A61P25/00
CPCC12N15/113A61P25/00C12N2310/532C12N2310/14C12N2310/113Y02A50/30
Inventor 巫荣华姚淳于彬刘梅刘炎
Owner NANTONG UNIVERSITY
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