A rapid method to screen putative mRNA targets of any known microRNA

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• 作者：Yujing Huang (1)
  Ying Qi (1)
  Qiang Ruan (1)
  Yanping Ma (1)
  Rong He (1)
  Yaohua Ji (1)
  Zhengrong Sun (1)
  
• 刊名：Virology Journal
• 出版年：2011
• 出版时间：December 2011
• 年：2011
• 卷：8
• 期：1
• 全文大小：1663KB
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• 作者单位：Yujing Huang (1)
  Ying Qi (1)
  Qiang Ruan (1)
  Yanping Ma (1)
  Rong He (1)
  Yaohua Ji (1)
  Zhengrong Sun (1)
  
  1. Virus Laboratory, the Affiliated Shengjing Hospital, China Medical University, 110004, Shenyang, Liaoning, PR China
  
• ISSN：1743-422X

文摘

Background microRNAs (miRNAs) are a group of regulatory RNAs that regulate gene expression by binding to specific sequences on target mRNAs. However, functional identification of mRNA targets is usually difficult and time consuming. Here we report hybrid-PCR as a new and rapid approach to screen putative mRNA targets in vitro. Results Fifteen putative target mRNAs for human cytomegalovirus (HCMV) miR-UL112-1, including previously confirmed HCMV IE72, were identified from mRNA-derived cDNAs using hybrid-PCR. Moreover, we randomly validated six different target candidates by luciferase reporter assays, and confirmed that their luciferase activities were down-regulated with co-transfection of HCMV miR-UL112-1. Conclusions Our study demonstrated that hybrid-PCR is an effective and rapid approach for screening putative miRNA targets, with much more advantage of simplicity, low cost, and ease of implementation.