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Circular RNA related to cardiac hypertrophy, expression vector and preparation method thereof

A technique for cardiac hypertrophy and expression vector, which is applied in the field of molecular biology and can solve the problem that the function and mechanism are not very clear.

Active Publication Date: 2022-01-04
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 analysis and detailed annotation of circRNAs were performed on cardiomyocyte ribosomal depletion RNA from human, mouse and rat hearts, as well as human embryonic stem cells (Qu S, Zhong Y, Shang R, et al. emerging landscape of circular RNA in lifeprocesses.RNA Biol.2017;14(8):992-9;Khan MA,Reckman YJ,Aufiero S,et al.RBM20Regulates 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 Mol Cell Cardiol.2016; 98:103-7.), found many in the heart Stably expressed circRNAs (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 cardiac hypertrophy, expression vector and preparation method thereof
  • Circular RNA related to cardiac hypertrophy, expression vector and preparation method thereof
  • Circular RNA related to cardiac hypertrophy, expression vector and preparation method thereof

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Embodiment

[0035] The concrete implementation process of the present invention is as follows:

[0036] 1. Construction of circular RNA chr4 133719537-133723051-adenovirus vector

[0037] 1.1 Design shRNA using Oligo Designer3.0

[0038] The loop structure in the ADV1-shRNA template uses TTCAAGAGA to avoid the formation of termination signals. AATTC was added to the 5' end of the sense strand template, which was complementary to the sticky end formed after EcoRI digestion; GATCC was added to the 5' end of the antisense strand template, which was complementary to the sticky end formed after BamHI digestion.

[0039] Sense strand: 5′- -(GN18)-( TTCAAGAGA )-(N18C)-TTTTTTG-3′(5′-AATTCGTTTCTCCGAACGTGTCACGTTTCAAGAGAACGTGACACGTTCGGAGAACTTTTTTG-3′)

[0040] Antisense strand: 3′-G(CN18)-( AAGTTCTCT )-(N18G)-AAAAAA -5'(5'-GATCCAAAAAAGTTCTCCGAACGTGTCACGTTCTCTTGAAACGTGACACGTTCGGAGAACG-3')

[0041] The relevant information of the target sequence numbered T6532 is as follows: figure 1 As sh...

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Abstract

The invention provides a circular RNA related to cardiac hypertrophy, an expression vector and a preparation method thereof. The circular RNA is discovered by sequencing a model mouse of cardiac hypertrophy, and plays a key role in the occurrence and development of cardiac 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, and its incidence has been increasing 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, myocardial hypertrophy is the most common pathological process in cardiovascular diseases, which can lead to abnormal hypertrophy of cardiomyocytes, hypertrophy and asymmetry of interventricular septum, narrowing of ventricular chambers or restriction of 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 receptor regulates cardiac hypertrophy and tr...

Claims

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Application Information

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