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. 2017 Nov 27;8:1546. doi: 10.3389/fimmu.2017.01546

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Copyright © 2017 He, Ma and Zhang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Mechanism of white spot syndrome virus (WSSV)-miR-N32 in virus infection. (A) Predicted target genes of WSSV-miR-N32. As predicted, the 3′ untranslated regions (3′UTRs) of the wsv459 and wsv322 genes were targeted by WSSV-miR-N32. The underline showed the seed sequence of WSSV-miR-N32. (B) The direct interactions between WSSV-miR-N32 and its target genes in insect cells. The insect High Five cells were co-transfected with WSSV-miR-N32 and enhanced green fluorescent protein (EGFP), EGFP-wsv459-3′UTR, EGFP-wsv322-3′UTR, EGFP-wsv459-3′UTR-mutation, or EGFP-wsv322-3′UTR-mutation. At 48 h after co-transfection, the fluorescence intensity of cells was examined. The sequences targeted by viral microRNA (miRNA) were underlined. (C) The effects of WSSV-miR-N32 on viral gene expressions. The relative fluorescence intensity of cells was determined. (D) The influence of WSSV-miR-N32 on viral gene expression in vivo. Shrimp were injected with WSSV and WSSV-miR-N32-mimic or anti-miRNA oligonucleotide (AMO)-WSSV-miR-N32. At 24 h after injection, the mRNA levels of wsv459 and wsv322 in shrimp hemolymph were examined with quantitative real-time polymerase chain reaction. Statistically significant differences between treatments were indicated by asterisks (*p < 0.05; **p < 0.01).

Mechanism of white spot syndrome virus (WSSV)-miR-N32 in virus infection. (A) Predicted target genes of WSSV-miR-N32. As predicted, the 3′ untranslated regions (3′UTRs) of the wsv459 and wsv322 genes were targeted by WSSV-miR-N32. The underline showed the seed sequence of WSSV-miR-N32. (B) The direct interactions between WSSV-miR-N32 and its target genes in insect cells. The insect High Five cells were co-transfected with WSSV-miR-N32 and enhanced green fluorescent protein (EGFP), EGFP-wsv459-3′UTR, EGFP-wsv322-3′UTR, EGFP-wsv459-3′UTR-mutation, or EGFP-wsv322-3′UTR-mutation. At 48 h after co-transfection, the fluorescence intensity of cells was examined. The sequences targeted by viral microRNA (miRNA) were underlined. (C) The effects of WSSV-miR-N32 on viral gene expressions. The relative fluorescence intensity of cells was determined. (D) The influence of WSSV-miR-N32 on viral gene expression in vivo. Shrimp were injected with WSSV and WSSV-miR-N32-mimic or anti-miRNA oligonucleotide (AMO)-WSSV-miR-N32. At 24 h after injection, the mRNA levels of wsv459 and wsv322 in shrimp hemolymph were examined with quantitative real-time polymerase chain reaction. Statistically significant differences between treatments were indicated by asterisks (*p < 0.05; **p < 0.01).