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

长链非编码RNA干扰素γ-反义1的功能和调控的表征

• 批准号: 10026713
• 负责人: Danielle Alexandria Chisolm
• 金额: --
• 依托单位: NATIONAL INST ARTH/MUSCLOSKEL/SKIN DIS
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10026713
• 关键词: ATAC-seq; Antigens; Architecture; Binding; Binding Proteins; Biological Assay; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CRISPR/Cas technology; Cell Line; ChIP-seq; Data; Development; Gene Expression; Gene Expression Regulation; Genome; Genomics; Glycolysis; Goals; Grant; Human; Human Genome; Immunize; Immunologic Memory; Interferon Type II; Knock-out; Lead; Lymphocytic choriomeningitis virus; Lymphoid Cell; Memory; Metabolic; Metabolism; Multiple Myeloma; Mus; Nutrient; Pathogenicity; Peptides; Plasma Cells; Play; Positioning Attribute; Production; Regulation; Regulatory Element; Research; Role; Sampling; Series; Supplementation; T-Lymphocyte; Techniques; Technology; Therapeutic Intervention; Untranslated RNA; Vaccinated; Virus; cell type; cytokine; experimental study; in vivo; inhibitor/antagonist; interleukin-22; new therapeutic target; next generation sequencing; novel; novel strategies; programs; response; tool; transcriptome sequencing

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.

项目总结/摘要 长链非编码RNA（lncRNA）在基因调控中起重要作用， 表达是一个快速发展的概念。lncRNA IFNG-AS 1存在于多种细胞因子中， 基因座，包括IFNG和IL 22。IFNG-AS 1已被证明在以下方面发挥重要作用： 在人和小鼠中正调节IFNγ表达。电流 关于Ifng-as 1作用的研究表明，它是Ifnγ的正调节剂，但 本授权中概述的初步数据显示了Ifng-as 1和 Ifng。这一发现表明这种lncRNA的替代调节。这项提案的目的是 了解IFNG-AS 1在不同细胞环境下的功能和调控。目标1 我们已经提出在不同的条件下定义IFNG-AS 1和基因组之间的关系， 使用HiChIRP（即，CD 4 + T细胞和浆细胞）的细胞设置和不同细胞状态。 我们还将确定哪些因子通过ATAC-seq与IFng-as 1差异结合， ChIP-seq.我们还将描述Ifng-as 1调节的代谢作用。 特别是通过使用代谢抑制剂和营养素的糖酵解和氨解 补充。在目标2中，我们将通过RAP-MS定义与IFng-as 1结合的蛋白质， 通过删除Ifng-as 1及其调控元件，定义了Ifng-as 1的功能， CRISPR/Cas9技术。最后，我们将试图确定IFNG-AS 1在体内的作用， ILC 3分化和记忆发育的环境。这项拟议研究的结果 将提供有关Ifng-as 1和Ifnγ调节的新机制信息 为治疗干预提供新的靶点。