High throughput functional characterization of lncRNAs in macrophage biology

巨噬细胞生物学中 lncRNA 的高通量功能表征

基本信息

批准号：
10874258
负责人：
Susan Carpenter
金额：
$ 8.2万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA CRUZ
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-07-31
项目状态：
未结题

项目摘要

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.

项目摘要在此提案中，我们结合了许多技术方法来提供一种新颖的方式在功能上表征复杂的基因网络，以识别功能调节巨噬细胞中的生物过程。深度测序技术的进步表明，大多数人类基因组积极转录为RNA。我们的实验室专注于表征最大的由名为长的非编码RNA（LNCRNA）的基因组产生的RNA组和它们相关的蛋白质结合伴侣。迄今为止，只有3％的lncrnas是在功能上验证。这个项目具有很高的创新性，因为我们将执行第一个系统的无偏屏幕，并创建第一个遗传相互作用图来识别与巨噬细胞内的生存力和功能有关的功能相关的lncRNA。在人和鼠标中使用我们新开发的记者细胞系，我们将能够快速筛选LNCRNA及其蛋白质结合伙伴的映射至关重要控制活力和炎症信号传导。我们还将获得有关的关键信息跨物种的LNCRNA功能保护。然后，我们将创建遗传鼠标模型证明这些基因及其调节网络在控制中的重要性体内败血症期间的免疫反应。这种方法将允许快速有意义以高效的方式获得的数据。实现雄心勃勃的目标提案将为我们提供有关涉及复杂途径的大量信息 lncrnas并洞悉其在有助于生存能力和功能方面的作用巨噬细胞。