Development of a micro-array to detect human and mouse microRNAs and characterization of expression in human organs
Development of a micro-array to detect human and mouse microRNAs and characterization of expression in human organs
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DOI：
10.1093/nar/gnh186
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发表时间：
2004-01-01
影响因子：
14.9
通讯作者：
Perera, RJ
中科院分区：
文献类型：
--
作者：
Sun, YQ;Koo, S;Perera, RJ
microRNA（miRNAs）被认为通过与互补的靶mRNA转录物杂交在发育和其他细胞过程中发挥重要作用。这导致杂交转录物的切割或翻译的负调控。关于miRNA表达的调控或模式知之甚少。高等真核生物中大量miRNA序列的预测存在使得单个miRNA分子本身的表达水平极有可能在调节多个细胞过程中发挥重要作用。因此，确定哺乳动物和其他高等真核生物中总体miRNA表达水平的模式对于帮助理解miRNA转录调控机制以及帮助鉴定miRNA调控的基因表达至关重要。在这里，我们描述了一种新的方法来检测全球加工的miRNA表达水平在高等真核生物，包括人类，小鼠和大鼠，通过使用高密度寡核苷酸阵列。阵列结果已经通过随后使用Northern印迹分析确认mir表达来验证。在来自不同来源的样品中已经检测到mir表达的主要差异，这表明高度调节的mir表达，以及个体miRNA转录物的特定基因调节功能。例如，与其他人体组织相比，发现五种不同的miRNA优先在人体肾脏中表达。肾脏特异性miRNA簇周围基因组序列的比较分析揭示了位于保守系统发育足迹中的特异性转录因子结合位点的存在，表明这些可能参与调节肾脏中的mir表达。
MicroRNAs (miRNAs) are believed to play important roles in developmental and other cellular processes by hybridizing to complementary target mRNA transcripts. This results in either cleavage of the hybridized transcript or negative regulation of translation. Little is known about the regulation or pattern of miRNA expression. The predicted presence of numerous miRNA sequences in higher eukaryotes makes it highly likely that the expression levels of individual miRNA molecules themselves should play an important role in regulating multiple cellular processes. Therefore, determining the pattern of global miRNA expression levels in mammals and other higher eukaryotes is essential to help understand both the mechanism of miRNA transcriptional regulation as well as to help identify miRNA regulated gene expression. Here, we describe a novel method to detect global processed miRNA expression levels in higher eukaryotes, including human, mouse and rats, by using a high-density oligonucleotide array. Array results have been validated by subsequent confirmation of mir expression using northern-blot analysis. Major differences in mir expression have been detected in samples from diverse sources, suggesting highly regulated mir expression, and specific gene regulatory functions for individual miRNA transcripts. For example, five different miRNAs were found to be preferentially expressed in human kidney compared with other human tissues. Comparative analysis of surrounding genomic sequences of the kidney-specific miRNA clusters revealed the occurrence of specific transcription factor binding sites located in conserved phylogenetic foot prints, suggesting that these may be involved in regulating mir expression in kidney.