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Circular RNA related to myocardial hypertrophy, expression vector and preparation method of expression vector

An expression vector and cardiac hypertrophy technology, applied in the field of molecular biology, can solve problems such as functions and mechanisms are not very clear

Pending Publication Date: 2020-10-09
THE SECOND HOSPITAL AFFILIATED TO SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Genome-wide profiling and detailed annotation of circRNAs in cardiomyocyte ribosome-depletion RNA from human, mouse, and rat hearts, as well as from human embryonic stem cells (Khan MA, Reckman YJ, Aufiero S, et al. RBM20 Regulates Circular RNA Production From the Titin Gene. Circ Res. 2016; 119(9):996-1003; Werfel S, Nothjunge S, Schwarzmayr T, et al. Characterization of circular RNAs in human, mouse and rat hearts. J MolCell Cardiol. 2016; 98:103-7; Tan WL, Lim BT, Anene-Nzelu CG, et al. A landscape of circular RNA expression in the human heart. Cardiovasc Res. 2017; 113(3):298-309), found many circRNAs stably expressed in the heart (Werfel S, Nothjunge S, Schwarzmayr T, et al. Characterization of circular RNAs in human, mouse and rat hearts. J Mol Cell Cardiol. 2016; 98:103-7; Tan WL, Lim BT, Anene- Nzelu CG, et al.A landscape of circular RNA expression in the human heart. Cardiovasc Res.2017; 113(3):298-309), but its function and mechanism are still not very clear

Method used

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  • Circular RNA related to myocardial hypertrophy, expression vector and preparation method of expression vector
  • Circular RNA related to myocardial hypertrophy, expression vector and preparation method of expression vector
  • Circular RNA related to myocardial hypertrophy, expression vector and preparation method of expression vector

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Embodiment

[0040] The specific implementation process of the present invention is as follows:

[0041] 1. Construction of Chr1 53256629-53282092 overexpression adenovirus vector

[0042] From the plasmid clone template containing the target gene, use the PCR method to fish for the target gene, if there is no template, use the method of whole gene synthesis to obtain the target gene. The target gene and the target vector were digested separately. After purification of the digested product, ligation is carried out, the ligated product is transformed into bacterial competent cells, and the grown clone is first identified by enzyme digestion to prove that the target gene has been directionally linked into the target vector. The positive clones are then sequenced, analyzed and compared, and the correct comparison is the successful construction of the target gene expression plasmid vector. The constructed recombinant vector is subjected to ultrapure extraction, the process is as follows fig...

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Abstract

The invention provides circular RNA related to myocardial hypertrophy, an expression vector and a preparation method of the expression vector. The circular RNA is found by sequencing a myocardial hypertrophy model mouse and plays a key role in the occurrence and development process of myocardial hypertrophy.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a circular RNA related to cardiac hypertrophy, an expression vector and a preparation method thereof. Background technique [0002] Cardiovascular disease is a major threat to human health with an increasing incidence in recent years (Molkentin JD, LuJR, Antos CL, et al. A calcineurin-dependent transcriptional pathway for cardiac hypertrophy. Cell 1998;93:215-28). Among them, cardiac hypertrophy is the most common pathological process in cardiovascular disease, which leads to abnormal hypertrophy of cardiomyocytes, hypertrophy and asymmetry of the ventricular septum, small intraventricular chambers, or restricted ventricular filling (Rockman HA, Koch WJ, Lefkowitz RJ. Seven-transmembrane-spanning receptors and heart function.Nature.2002;415:206-12;Everett AD,Tufro-McReddie A,Fisher A,etal.Angiotensin receptors regulates cardiac hypertrophy and transforming growthfactor-be...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/113C12N15/861C12N15/66
CPCC12Q1/6883C12N15/86C12N15/66C12N15/113C12Q2600/158C12Q2600/178C12N2710/10043C12N2310/532Y02A50/30
Inventor 周祥王文静陈丽莉
Owner THE SECOND HOSPITAL AFFILIATED TO SUZHOU UNIV