A precise and efficient circular RNA synthesis system based on a ribozyme derived from <i>Tetrahymena thermophila</i>

doi:10.1093/nar/gkad554

Jingyi Cui, Lanxin Zhang, Zaifeng Zhang, Xuanmei Luo, Ye Liu, Chang Li, Wei Huang, Lihui Zou, Xue Yu, Fei Xiao

A precise and efficient circular RNA synthesis system based on a ribozyme derived from Tetrahymena thermophila

Genetics

Abstract Classic strategies for circular RNA (circRNA) preparation always introduce large numbers of linear transcripts or extra nucleotides to the circularized product. In this study, we aimed to develop an efficient system for circRNA preparation based on a self-splicing ribozyme derived from an optimized Tetrahymena thermophila group Ⅰ intron. The target RNA sequence was inserted downstream of the ribozyme and a complementary antisense region was added upstream of the ribozyme to assist cyclization. Then, we compared the circularization efficiency of ribozyme or flanking intronic complementary sequence (ICS)-mediated methods through the DNMT1, CDR1as, FOXO3, and HIPK3 genes and found that the efficiency of our system was remarkably higher than that of flanking ICS-mediated method. Consequently, the circularized products mediated by ribozyme are not introduced with additional nucleotides. Meanwhile, the overexpressed circFOXO3 maintained its biological functions in regulating cell proliferation, migration, and apoptosis. Finally, a ribozyme-based circular mRNA expression system was demonstrated with a split green fluorescent protein (GFP) using an optimized Coxsackievirus B3 (CVB3) internal ribosome entry site (IRES) sequence, and this system achieved successful translation of circularized mRNA. Therefore, this novel, convenient, and rapid engineering RNA circularization system can be applied for the functional study and large-scale preparation of circular RNA in the future.

Need a simple solution for managing your BibTeX entries? Explore CiteDrive!

Web-based, modern reference management
Collaborate and share with fellow researchers
Integration with Overleaf
Comprehensive BibTeX/BibLaTeX support
Save articles and websites directly from your browser
Search for new articles from a database of tens of millions of references

Try out CiteDrive

A precise and efficient circular RNA synthesis system based on a ribozyme derived from Tetrahymena thermophila

Need a simple solution for managing your BibTeX entries? Explore CiteDrive!

More from our Archive

CRISPR‐Cas9 screening identified lethal genes enriched in necroptosis pathway and of prognosis significance in osteosarcoma

Identification of the shared gene signatures and molecular pathways in systemic lupus erythematosus and diffuse large B‐cell lymphoma

Nanoparticles‐induced potential toxicity on human health: Applications, toxicity mechanisms, and evaluation models

The reconfiguration pattern of individual brain metabolic connectome for Parkinson's disease identification

Omics-based deep learning approaches for lung cancer decision-making and therapeutics development

Multi-Omics Reveals the Impact of Exogenous Short-Chain Fatty Acid Infusion on Rumen Homeostasis: Insights into Crosstalk between the Microbiome and the Epithelium in a Goat Model

Spinal Muscular Atrophy Treatment in Patients Identified by Newborn Screening—A Systematic Review

CDK8 and CDK19: positive regulators of signal-induced transcription and negative regulators of Mediator complex proteins

TLK1-mediated RAD54 phosphorylation spatio-temporally regulates Homologous Recombination Repair

<scp>RBOHF</scp> activates stomatal immunity by modulating both reactive oxygen species and apoplastic <scp>pH</scp> dynamics in Arabidopsis