MicroRNA function in murine cytomegalovirus

MicroRNA在小鼠巨细胞病毒中的功能

• 批准号: BB/G01552X/1
• 负责人: Amy Buck
• 金额: $ 42.24万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2009
• 资助国家: 英国
• 起止时间: 2009 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FG01552X%2F1
• 关键词: MicroRNA; function; murine; cytomegalovirus

The broad goal of this project is to determine the mechanism by which an animal virus utilizes and controls microRNA (miRNA) expression in order to establish an infection and persist in its host. MiRNAs are a class of small RNA molecules (~ 22 nucleotides long), discovered in the last decade, that regulate gene expression by targeting specific messenger RNA transcripts for degradation or inhibited translation. It is estimated that 30% of genes in humans could be regulated by miRNAs, implicating a fundamental role of these molecules in modulating the transcriptome. Viral-encoded miRNAs have been discovered recently in a number of different viruses, including all three herpesvirus subfamilies. Recent reports have demonstrated that viral miRNAs can target host genes involved in the immune response. This class of molecules, therefore, represents an important new class of potential drug targets. To date, no viral miRNA has been examined in vivo. Understanding the functional role of miRNAs in an intact physiological system is of vital importance; an in vivo analysis considers that both viral and cellular gene expression can be influenced by cell-context as well as the extent of the anti-viral immune response. We will examine viral miRNA function in murine cytomegalovirus (CMV). Cytomegalovirus, a member of the betaherpesvirus family, is a ubiquitous virus that is a major cause of morbidity in the clinical setting. CMV is highly species-specific; however, the pathogenesis of MCMV in mice is remarkably similar to that of HCMV in humans. Murine CMV is therefore an established model for studying human CMV and the MCMV genome can be modified to generate miRNA deletion mutants using bacterial artificial chromosome (BAC) technology. We will use both MCMV miRNA deletion mutants and antisense technology to examine viral miRNA function in vivo. Microarray technology will be used to examine the host genes that are targeted by MCMV miRNAs. This work is highly relevant to future analysis of miRNA inhibitors as anti-viral therapeutics. Finally we will establish a system for examining viral miRNA biogenesis in vitro. This is required to gain an in-depth understanding of how the virus controls and exploits this class of molecules and should complement the in vivo analysis. The tools and knowledge resulting from this work will be widely applicable in other viral systems. MiRNAs have also been shown to play a role in cancer formation and cardiovascular and metabolic diseases; expanding the understanding of miRNA function and mechanism, therefore, is of paramount importance to human health.

该项目的广泛目标是确定动物病毒利用和控制microRNA（miRNA）表达以建立感染并持续在其宿主中的机制。 miRNA是在过去十年中发现的一类小RNA分子（〜22个核苷酸长），它们通过靶向特定的使者RNA转录本来调节基因表达，以降解或抑制翻译。据估计，人类中有30％的基因可以受miRNA调节，这暗示了这些分子在调节转录组中的基本作用。病毒编码的miRNA最近在许多不同的病毒中发现，包括所有三种疱疹病毒亚家族。最近的报告表明，病毒miRNA可以靶向涉及免疫反应的宿主基因。因此，这类分子代表了一类重要的潜在药物靶标。迄今为止，尚未在体内检查病毒miRNA。了解miRNA在完整生理系统中的功能作用至关重要。体内分析认为，病毒和细胞基因表达都可以受细胞膜的影响以及抗病毒免疫反应的程度。我们将检查鼠巨细胞病毒（CMV）中的病毒miRNA功能。 Betaherpesvirus家族的成员巨细胞病毒是一种普遍存在的病毒，是临床环境中发病率的主要原因。 CMV是高度物种特异性的；然而，小鼠中MCMV的发病机理与人类的HCMV非常相似。因此，鼠CMV是研究人CMV的既定模型，并且可以使用细菌性人工染色体（BAC）技术来修改MCMV基因组以生成miRNA缺失突变体。我们将使用MCMV miRNA缺失突变体和反义技术来检查体内病毒miRNA功能。微阵列技术将用于检查MCMV miRNA针对的宿主基因。这项工作与将miRNA抑制剂作为抗病毒疗法的未来分析高度相关。最后，我们将建立一个在体外检查病毒miRNA生物发生的系统。这是对病毒如何控制和利用这类分子的深入了解，并应补充体内分析所需的。这项工作产生的工具和知识将广泛适用于其他病毒系统。还显示miRNA在癌症形成以及心血管和代谢疾病中发挥作用。因此，扩展对miRNA功能和机制的理解对人类健康至关重要。

项目成果

Host gene targets for novel influenza therapies elucidated by high-throughput RNA interference screens.

• DOI: 10.1096/fj.11-193466
• 发表时间: 2012-04
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Meliopoulos VA;Andersen LE;Birrer KF;Simpson KJ;Lowenthal JW;Bean AG;Stambas J;Stewart CR;Tompkins SM;van Beusechem VW;Fraser I;Mhlanga M;Barichievy S;Smith Q;Leake D;Karpilow J;Buck A;Jona G;Tripp RA
• 通讯作者: Tripp RA