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Circular RNA circTTC3 overexpression adeno-associated virus vector, adeno-associated virus and application of adeno-associated virus

A viral vector, overexpression technology, applied in the field of biomedicine, can solve the problem of lack of circular RNA diagnosis, prevention and treatment

Active Publication Date: 2022-05-20
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there is still a lack of reports on the diagnosis, prevention and treatment of circular RNAs for myocardial ischemia-reperfusion injury.

Method used

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  • Circular RNA circTTC3 overexpression adeno-associated virus vector, adeno-associated virus and application of adeno-associated virus
  • Circular RNA circTTC3 overexpression adeno-associated virus vector, adeno-associated virus and application of adeno-associated virus
  • Circular RNA circTTC3 overexpression adeno-associated virus vector, adeno-associated virus and application of adeno-associated virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051]1. Mouse model of myocardial ischemia-reperfusion injury

[0052] Mice were anesthetized by intraperitoneal injection of 1 mg / 10 g body weight of pentobarbital sodium. Remove hair with depilatory cream. The mice were fixed on a 37°C warming pad. An incision is made along the trachea, and the muscles and tissues covering the trachea are carefully separated to expose the trachea. An endotracheal tube was inserted between the subglottic tracheal cartilages and fixed. The assisted breathing rate was 120 times / min, and the tidal volume was 2 mL. Make an incision of about 1 cm at about 0.5 cm under the armpit, and cut the skin, pectoralis major to intercostal muscles layer by layer to expose. Insert blunt curved ophthalmic forceps between the third and fourth ribs, and bluntly separate the intercostal muscles to expose the heart. About 1 / 3 of the line from the left atrial appendage to the apex was ligated with a low-elasticity 7-0 suture with a slipknot. The whitening bel...

Embodiment 2

[0072] 1. Isolation and Culture of Primary Rat Cardiomyocytes

[0073] 75% alcohol sterilized the chest of newborn rats, took out the rat heart under aseptic conditions, dissected and separated the ventricle tissue, and placed the ventricle tissue in iced 1×ADS buffer solution (preparation of 1L 10×ADS buffer solution: 68g sodium chloride, 47.6 g 4-hydroxyethylpiperazineethanesulfonic acid, 1.38g sodium dihydrogen phosphate, 6g glucose, 4g potassium chloride, 2.05g magnesium sulfate, rinse with sterilized water to 1L), put the tissue in a sterile glass Cut into pieces in the bottle, add 20mL trypsin collagenase digestion solution for digestion. Put the digestion flask in a shaker (37°C, 90-120rpm), digest for 10 minutes each time, then transfer the cell suspension to a 50mL centrifuge tube, add 1 / 5 volume of horse serum to stop the digestion, and centrifuge at 1,000rmp for 5min. Discard the cell supernatant, resuspend, repeat the above digestion steps until it becomes floccul...

Embodiment 3

[0077] 1. Isolation and Culture of Primary Rat Cardiomyocytes

[0078] 75% alcohol sterilized the chest of newborn rats, took out the rat heart under aseptic conditions, dissected and separated the ventricle tissue, and placed the ventricle tissue in iced 1×ADS buffer solution (preparation of 1L 10×ADS buffer solution: 68g sodium chloride, 47.6 g 4-hydroxyethylpiperazineethanesulfonic acid, 1.38g sodium dihydrogen phosphate, 6g glucose, 4g potassium chloride, 2.05g magnesium sulfate, rinse with sterilized water to 1L), put the tissue in a sterile glass Cut into pieces in the bottle, add 20mL trypsin collagenase digestion solution for digestion. Put the digestion flask in a shaker (37°C, 90-120rpm), digest for 10 minutes each time, then transfer the cell suspension to a 50mL centrifuge tube, add 1 / 5 volume of horse serum to stop the digestion, and centrifuge at 1,000rmp for 5min. Discard the cell supernatant, resuspend, repeat the above digestion steps until it becomes floccul...

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Abstract

The invention provides a circular RNAcircTTC3 overexpression adeno-associated virus vector, an adeno-associated virus and application of the adeno-associated virus, and belongs to the technical field of biological medicine. The cyclic RNA-circTTC3 delivered by using the AAV9 virus vector can be used for preparing a medicine for preventing and / or treating myocardial ischemia-reperfusion injury (IRI). The gene sequence capable of transcribing the circular RNA-circTTC3 is delivered to the heart tissue based on the AAV virus, and is stably and highly expressed in the heart tissue, the circTTC3 is up-regulated to generate a heart protection effect, and the myocardial infarction area is effectively reduced, so that the effect of protecting myocardial infarction is achieved. According to the application disclosed by the invention, circTTC3 is over-expressed in primary myocardial cells of newborn rats, and the circTTC3 is found to be capable of remarkably promoting proliferation and growth of the primary myocardial cells of the newborn rats.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a circular RNA circTTC3 overexpression adeno-associated virus vector, an adeno-associated virus and applications thereof. Background technique [0002] Acute myocardial infarction is one of the leading causes of cardiovascular death and disability. For patients with acute myocardial infarction, myocardial ischemia reperfusion therapy, which is commonly used clinically, can reduce the acute phase death caused by myocardial infarction to a certain extent, but the myocardial damage caused by itself and the adverse promotion effect on ventricular remodeling Seriously affect the prognosis of patients with myocardial infarction, this process and phenomenon is called myocardial ischemia-reperfusion injury. Therefore, the study of effective against myocardial ischemia-reperfusion injury has been widely concerned. At present, the prevention and treatment of myocardial is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/864C12N15/113A61K31/7105A61P9/10C12Q1/6883C12N15/11
CPCC12N15/86C12N15/113A61K31/7105A61P9/10C12Q1/6883C12N2750/14143C12N2800/107C12N2310/532C12N2320/32C12Q2600/158C12Q2600/178
Inventor 肖俊杰王丽君陈奕文吴心盈杨子江
Owner SHANGHAI UNIV
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