Characterization of the function and regulation of long noncoding RNA Interferon gamma- anti-sense 1

项目摘要/摘要 认为长非编码核糖核酸(LncRNA)在基因调控中起重要作用的观点 表达是一个快速发展的概念。IncRNA IFNG-AS1存在于多种细胞因子中 包括IFNG和IL22的基因座。IFNG-AS1已被证明在 正向调节干扰素γ在人和小鼠体内的表达。当前 关于IFNG-AS1作用的研究表明，它是干扰素γ的正向调节因子，但 本授权中概述的初步数据显示了IFNG-AS1和IFNG-AS1不一致表达的示例 IFNG。这一发现提出了对这种lncRNA的另一种调节。这项提议的目标是 了解IFNG-AS1在不同细胞环境下的功能和调控。在目标1中 我们已经提出了在不同的条件下定义IFNG-AS1与基因组的关系 使用HiChIRP的细胞设置和不同的细胞状态(即CD4+T细胞和浆细胞)。 我们还将通过atac-seq确定哪些因子与IFNG-AS1和 芯片顺序我们还将描述IFNG-AS1在体内的代谢作用。 通过使用代谢抑制剂和营养素进行特殊的糖酵解和谷氨酰胺分解 补充。在目标2中，我们将通过RAP-MS定义与IFNG-AS1结合的蛋白质，并 定义了IFNG-AS1的功能，删除了IFNG-AS1及其调节元件 CRISPR/CAS9技术。最后，我们将寻求确定IFNG-AS1在体内的作用 ILC3分化和记忆力发展的设置。这项拟议研究的发现 将提供有关干扰素-as1和干扰素γ调控的新的机制信息 为治疗干预提供新的靶点。

Project Summary/Abstract The idea that long noncoding RNAs (lncRNA) play an important role in regulating gene expression is a quickly growing concept. The lncRNA IFNG-AS1 is found in a multi-cytokine locus which includes IFNG and IL22. IFNG-AS1 has been shown to play an important role for positively regulating the expression of IFNγ expression in both human and mice. Current research concerning the role for Ifng-as1 suggests its role as a positive regulator of Ifnγ, but preliminary data outlined in this grant shows examples of discordant expression of Ifng-as1 and Ifng. This finding suggests an alternative regulation of this lncRNA. The goal of this proposal is to understand the function and regulation of Ifng-as1 under different cellular settings. In Aim 1 we have proposed to define the relationship between Ifng-as1 and the genome under different cellular settings and different cellular states using HiChIRP (i.e. CD4+ T cells and plasma cells). We will also determine what factors are differentially bound to Ifng-as1 through ATAC-seq and ChIP-seq. We will additionally describe a metabolic role for the regulation of Ifng-as1 in particular glycolysis and glutaminolysis through the use of metabolic inhibitors and nutrient supplementation. In Aim 2 we will define proteins bound to Ifng-as1 through RAP-MS and defined the function of Ifng-as1 by deleting Ifng-as1 and its regulatory elements by using CRISPR/Cas9 technology. Finally, we will seek to determine a role for Ifng-as1 in an in vivo setting for ILC3 differentiation and memory development. The findings from this proposed study will provide novel mechanistic information concerning the regulation of Ifng-as1 and Ifnγ providing novel targets for therapeutic intervention.