High throughput functional characterization of lncRNAs in macrophage biology

项目摘要 在这个提议中，我们联合收割机结合了许多技术方法，提供了一种新的方式 功能上表征复杂的基因网络，以确定那些功能， 调节巨噬细胞中的生物过程。 深度测序技术的进展表明，大多数人类 基因组被主动转录成RNA。我们的实验室专注于表征 由基因组产生的一组RNA，称为长链非编码RNA（lncRNA）， 它们的相关蛋白结合伴侣。到目前为止，只有3%的lncRNA被 功能验证。这个项目是高度创新的，因为我们将执行第一个 系统的无偏筛选，并创建第一个遗传相互作用图，以确定 功能相关的lncRNA参与巨噬细胞内的活力和功能。 使用我们新开发的人类和小鼠报告细胞系，我们将能够 快速筛选和定位lncRNA及其蛋白结合伴侣， 控制存活力和炎症信号。我们还将获得有关以下方面的重要信息： 跨物种lncRNA的功能保守性。然后我们将创造一个基因小鼠 模型来证明这些基因及其调控网络在控制 体内脓毒症期间的免疫应答。这种方法将允许快速有意义的 以高效的方式获得数据。实现这一宏伟目标 该提案将为我们提供有关复杂途径的丰富信息， lncRNA，并深入了解它们在促进细胞活力和功能方面的作用。 巨噬细胞

Project Summary In this proposal we combine a number of technological approaches to provide a novel way to functionally characterize complex gene networks to identify those that function to regulate biological processes in macrophages. Advances in deep sequencing technologies have revealed that the majority of the human genome is actively transcribed into RNA. Our lab is focused on characterizing the largest group of RNA produced from the genome named long noncoding RNA (lncRNAs) and their associated protein binding partners. To date only 3% of lncRNAs have been functionally validated. This project is highly innovative as we will perform the first systematic unbiased screens and create the first genetic interaction maps to identify functionally relevant lncRNAs involved in viability and functions within macrophages. Using our newly developed reporter cell lines in both human and mouse we will be able to rapidly screen and map for lncRNAs and their protein binding partners that are critical for controlling viability and inflammatory signaling. We will also obtain crucial information on functional conservation of lncRNAs across species. We will then create genetic mouse models to prove the importance of these genes and their regulatory networks in controlling immune responses during sepsis in vivo. This approach will allow for rapid meaningful data to be obtained in a highly efficient manner. Accomplishing the ambitious goals of this proposal will provide us with a wealth of information on the complex pathways involving lncRNAs and gain insights into their roles in contributing to viability and functions of macrophages.