TAILIEUCHUNG - Báo cáo y học: " A rapid method to screen putative mRNA targets of any known microRNA"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: A rapid method to screen putative mRNA targets of any known microRNA | Huang et al. Virology Journal 2011 8 8 http content 8 1 8 J VIROLOGY JOURNAL METHODOLOGY Open Access A rapid method to screen putative mRNA targets of any known microRNA Yujing Huang Ying Qi Qiang Ruan Yanping Ma Rong He Yaohua Ji Zhengrong Sun Abstract 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. Background MicroRNAs miRNAs are the most studied non-coding RNAs in recent years. miRNAs are 17- to 30-nucleotide RNAs that are ubiquitously expressed in plants and animals. They regulate gene expression at the posttran-scriptional level 1 2 and act as key regulators in diverse regulatory pathways including early development cell differentiation cell proliferation metabolism and apoptosis 3-6 . miRNAs binding to target mRNAs often leads to blockade of translation or degradation of the target mRNAs. Identification of target mRNAs is essential for understanding the biological functions of miRNAs. miRNAs from plants induce direct cleavage and degradation by binding to the target sequences with perfect base pairing. Targets of mammalian miRNAs are often difficult to predict because few of them match to their .

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