High throughput functional characterization of lncRNAs in macrophage biology

项目摘要 在这个提案中，我们结合了许多技术方法，以提供一种新的方式 从功能上描述复杂的基因网络，以确定那些对 调节巨噬细胞的生物学过程。 深度测序技术的进步表明，大多数人类 基因组被活跃地转录成RNA。我们的实验室专注于描述最大的 从基因组中产生的一组RNA，称为长非编码RNA(LncRNAs)和 它们的相关蛋白质结合伙伴。到目前为止，只有3%的lncRNA是 在功能上经过验证。这个项目是高度创新的，因为我们将执行第一个 系统化的无偏见筛选和创建第一个遗传相互作用图谱来识别 与巨噬细胞内活性和功能相关的功能相关的lncRNAs。 利用我们新开发的人类和小鼠报告细胞系，我们将能够 快速筛选和绘制lncRNAs及其蛋白质结合伙伴的图谱 控制生存和炎症信号。我们还将获得有关以下方面的关键信息 跨物种的lncRNAs的功能保守。然后我们将创造出遗传老鼠 证明这些基因及其调控网络在控制中的重要性的模型 体内脓毒症时的免疫反应。这种方法将允许快速有意义的 以高效的方式获取数据。实现这个雄心勃勃的目标 提案将为我们提供有关复杂途径的丰富信息 并深入了解它们在促进人类生存能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.