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Figure 5 | Virology Journal

Figure 5

From: Identification and validation of a novel microRNA-like molecule derived from a cytoplasmic RNA virus antigenome by bioinformatics and experimental approaches

Figure 5

Detection of hav-miR-N1-3p from plasmid-expressed precursor miRNA (pre-miRNA). (A) Construction of miRNA expression plasmid. Each pre-miRNA sequence was cloned into a pcDNA6.2-GW/EmGFP-miR expression vector. The vector with scrambled sequences was used as a negative control. (B) KMB17 and HEK293T cells were transfected with miRNA expression plasmid, and strong green fluorescence was observed in both transfected cell lines (100 ×). (C) Detection of mature miRNA hav-miR-N1-3p expression. Small RNA was harvested at 24 hours post-transfection, and mature hav-miR-N1-3p was detected by stem-loop RT-PCR in both transfected cells. (D) Plasmid-expressed hav-miR-N1-3p was reduced in Dicer-deficient KMB17 and HEK293T cells compared with Dicer-non-deficient negative control cells. Significant difference in expression levels were detected by Student’s t-test (*P < 0.05 and **P < 0.01).

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Virology Journal

ISSN: 1743-422X