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A universal approach to investigate circRNA protein coding function.

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机构: [1]Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Strasse 9b, 50931, Cologne, Germany [2]Medical Faculty, Core Facility Transgenic Animal and Genetic Engineering Models (TRAM), University of Münster, Von-Esmarch-Str.56, D-48149, Münster, Germany [3]Institute of Experimental Pathology, Centre for Molecular Biologyof Inflammation (ZMBE), University of Münster, Von-Esmarch-Str.56] D-48149, Münster, Germany [4]Brandenburg Medical School (MHB), Fehrbelliner Strasse 38, D-16816, Neuruppin, Germany [5]Institute of Medical Biochemistry,Centre for Molecular Biology of Inflammation (ZMBE), University of Münster, Von-Esmarch-Strasse 56, D-48149,Münster, Germany [6]Institutes for Systems Genetics, West China Hospital, Sichuan University, Chengdu, 610041,China
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摘要:
Circular RNAs (circRNAs) are an emerging class of RNA molecules that have been linked to human diseases and important regulatory pathways. Their functional roles are still under investigation, often hampered by inefficient circRNA formation in and ex vivo. We generated an intron-mediated enhancement (IME) system that-in comparison to previously published methods-increases circRNA formation up to 5-fold. This strategy also revealed previously undetected translation of circRNA, e.g., circRtn4. Substantiated by Western blots and mass spectrometry we showed that in mammalian cells, translation of circRtn4 containing a potential "infinite" circular reading frame resulted in "monomers" and extended proteins, presumably "multimer" tandem repeats. In order to achieve high levels of circRNA formation and translation of other natural or recombinant circRNAs, we constructed a versatile circRNA expression vector-pCircRNA-DMo. We demonstrated the general applicability of this method by efficiently generating two additional circRNAs exhibiting high expression levels. The circRNA expression vector will be an important tool to investigate different aspects of circRNA biogenesis and to gain insights into mechanisms of circular RNA translation.

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出版当年[2019]版:
大类 | 3 区 综合性期刊
小类 | 3 区 综合性期刊
最新[2023]版:
大类 | 2 区 综合性期刊
小类 | 2 区 综合性期刊
第一作者:
第一作者机构: [1]Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Strasse 9b, 50931, Cologne, Germany [2]Medical Faculty, Core Facility Transgenic Animal and Genetic Engineering Models (TRAM), University of Münster, Von-Esmarch-Str.56, D-48149, Münster, Germany
通讯作者:
通讯机构: [1]Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Strasse 9b, 50931, Cologne, Germany [2]Medical Faculty, Core Facility Transgenic Animal and Genetic Engineering Models (TRAM), University of Münster, Von-Esmarch-Str.56, D-48149, Münster, Germany
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