Establishment of Methods for Studying Circular RNAs’ Translational Function
LI Zhe, XIAO Bingxiu*
The discovery of the translation function of circRNAs (circular RNAs) has changed the traditional understanding of circRNAs as a non-coding RNAs and has become an important direction in the functional research field of circRNAs. The purpose of this paper is to establish a method for studying the translation function of circRNAs. In this study, based on the results of circRNA microarray detection in cancer tissue and plasma of patients with gastric cancer, synchronous changes of circRNAs were screened by ORF finder (open reading frame finder) and CPAT (coding-potential assessment tool), to detect the feasibility of ORF and translation of the above circRNAs, respectively. IRES finder (internal ribosome entry site finder) and circRNADb database were used to analyze whether there were IRES (internal ribosome entry site) in circRNAs, and then M6AMRFS (robust prediction of N6-methyladenosine sites with sequence-based features in multiple species) tool and SRAMP (sequence-based RNA adenosine methylation site predictor) tool were used to screen whether circRNAs contained M6A modification sites. Finally, the circRNAs with translation function were screened out by intersection analysis. The results show that seven kinds of circRNAs may have translation function, among which hsa_circ_0001479 has both IRES and M6A-mediated translation function; hsa_circ_0001278, hsa_circ_0001439 and hsa_circ_0000220 may mediate translation through IRES, while hsa_circ_0001017, hsa_circ_0008351 and hsa_circ_0004406 may mediate translation through M6A. This study provides a reference for exploring whether circRNAs has translation function. The feasibility of the selected analysis method is verified by constructing overexpression plasmid and Western blot. The results show that a set of feasible methods for the study of the translation function of circRNAs has been successfully established.
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