A rapid method to screen putative mRNA targets of any known microRNA.
A rapid method to screen putative mRNA targets of any known microRNA.
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一种快速筛选任何已知 microRNA 的假定 mRNA 靶标的方法
DOI：
10.1186/1743-422x-8-8
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发表时间：
2011-01-11
期刊：
影响因子：
4.8
通讯作者：
Sun Z
中科院分区：
文献类型：
--
作者：
Huang Y;Qi Y;Ruan Q;Ma Y;He R;Ji Y;Sun Z
研究背景microRNAs（miRNAs）是一类通过与靶mRNA上的特定序列结合来调控基因表达的调控RNA。然而，mRNA靶标的功能鉴定通常是困难且耗时的。在这里，我们报告杂交-PCR作为一种新的和快速的方法来筛选推定的mRNA targets in vitro. Results 15推定的靶mRNA的人巨细胞病毒（HCMV）的miR-UL 112 -1，包括先前确认的HCMV IE72，从mRNA衍生的cDNA使用杂交-PCR。同时，我们还对6个候选靶点进行了荧光素酶报告基因检测，并证实了它们的荧光素酶活性在HCMV miR-UL 112 - 1共转染后均被下调。结论杂交PCR技术是一种快速有效的筛选miRNA靶点的方法，具有操作简单、成本低、易于实现等优点。
BackgroundmicroRNAs (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.ResultsFifteen 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.ConclusionsOur 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.