Characterization analysis and overexpressed vector construction of the circular RNA hsa_circ_0082626 derived from the antiviral gene ZC3HAV1 / 医学研究生学报

ABSTRACT

Objective Circular RNA is a research hotspot of non-coding RNAs. The purpose of this study was to investigate the basic characteristics and expression effect of the overexpression vectors of circular RNA hsa_circ_0082626 transcribed from antiviral gene ZC3HAV1. Methods The basic characteristics of hsa_circ_0082626 were studied by the verification of reverse cleavage site, RNase digestion assay and intracellular distribution location with extraction of cytoplasmic and nuclear RNA. In the RNase digestion assay, samples were divided into RNase R treatment group and control group. RNase. 20 U (2 U/μg) of RNase R was added to the R treatment group, and the control group was replaced with an equivalent amount of ddH2O to detect changes in expression levels after RNase treatment. The cells were divided into 2 groups overexpression group and negative control group. At 24, 48 and 72 h after transfection, cells were collected to detect the expression of circular RNA by RT-qPCR. Results Compared with the control group, the expression of ZC3HAV1, CDR1as and GAPDH in the RNase R treatment group was increased (0.144±0.002 vs 1.000±0.016, 0.772±0.058 vs 1.000±0.122, 0.077±0.009 vs 1.000±0.164, P<0.05). Hsa_circ_0082626 could resist the treatment of RNase R and was mainly distributed in cytoplasm. The expression level of hsa_circ_0082626 in the overexpression group was significantly higher than that in the negative control group, and the expression level was the highest at 48 h after transfection. Conclusion The characteristics of hsa_circ_0082626 reverse cleavage site, RNase resistance and expression in cells were successfully analyzed, which proved that hsa_circ_0082626 does have a circular structure. The overexpression vector of hsa_circ_0082626 was successfully constructed to provide an experimental basis for the biological function and mechanism of RNA hsa_circ_0082626.