Yuan Ji-Min, Chen Yong-Shun, He Jian, Weng Shao-Ping, Guo Chang-Jun, He Jian-Guo
Guangdong Provincial Key Laboratory of Marine Resources and Coastal Engineering/South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation Center, School of Marine, Sun Yat-sen University, 135 Xingang Road West, Guangzhou, 510275, PR China.
State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, 135 Xingang Road West, Guangzhou, 510275, PR China.
Virol J. 2016 Apr 30;13:73. doi: 10.1186/s12985-016-0530-6.
Tiger frog virus (TFV), dsDNA virus of the genus Ranavirus and family Iridoviridae, causes a high mortality of tiger frog tadpoles cultured in Southern China. MicroRNAs (miRNAs) have been identified in many viruses especially DNA viruses such as Singapore Grouper Iridoviruses (SGIV). MicroRNAs play important roles in regulating gene expression for virus subsistence in host. Considering that TFV infects cells of different species under laboratory conditions, we aim to identify the specific and essential miRNAs expressed in ZF4 and HepG2 cells.
We identified and predicted novel viral miRNAs in TFV-infected ZF4 and HepG2 cells by deep sequencing and software prediction. Then, we verified and described the expression patterns of TFV-encoded miRNAs by using qRT-PCR and Northern blot.
Deep sequencing predicted 24 novel TFV-encoded miRNAs, and qRT-PCR verified 19 and 23 miRNAs in TFV-infected ZF4 (Group Z) and HepG2 (Group H) cells, respectively. Northern blot was performed to validate eight and five TFV-encoded miRNAs in Groups H and Z, respectively. We compared the expression of TFV-encoded miRNAs from two groups and defined TFV-miR-11 as the essential viral miRNA and TFV-miR-13 and TFV-miR-14 as the specific miRNAs that contribute to HepG2 cell infection.
We identified novel viral miRNAs and compared their expression in two host cells. The results of this study provide novel insights into the role of viral miRNAs in cross-species infection in vitro.
虎纹蛙病毒（TFV）是虹彩病毒科蛙病毒属的双链DNA病毒，可导致中国南方养殖的虎纹蛙蝌蚪高死亡率。微小RNA（miRNA）已在许多病毒尤其是DNA病毒如新加坡石斑鱼虹彩病毒（SGIV）中被鉴定出来。微小RNA在调节病毒在宿主中生存所需的基因表达方面发挥重要作用。鉴于TFV在实验室条件下可感染不同物种的细胞，我们旨在鉴定在ZF4和HepG2细胞中表达的特异性和必需的miRNA。
我们通过深度测序和软件预测在TFV感染的ZF4和HepG2细胞中鉴定并预测新型病毒miRNA。然后，我们使用qRT-PCR和Northern印迹验证并描述TFV编码的miRNA的表达模式。
深度测序预测了24种新型TFV编码的miRNA，qRT-PCR分别在TFV感染的ZF4（Z组）和HepG2（H组）细胞中验证了19种和23种miRNA。Northern印迹分别对H组和Z组中的8种和5种TFV编码的miRNA进行了验证。我们比较了两组中TFV编码的miRNA的表达，并将TFV-miR-11定义为必需的病毒miRNA，将TFV-miR-13和TFV-miR-14定义为有助于HepG2细胞感染的特异性miRNA。
我们鉴定了新型病毒miRNA并比较了它们在两种宿主细胞中的表达。本研究结果为病毒miRNA在体外跨物种感染中的作用提供了新的见解。
