Determining the factors that control dose-dependent splicing regulation by a master regulator

确定主调节器控制剂量依赖性剪接调节的因素

• 批准号: 9383785
• 负责人: Andrew Berglund
• 金额: $ 42.11万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-04 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9383785
• 关键词: Address; Affect; Alternative Splicing; Architecture; Behavior; Binding; Binding Sites; Biological; Cell model; Cell physiology; Code; Cooperative Behavior; Data; Development; Disease; Dose; Elements; Event; Exhibits; Exons; Explosion; Heart Diseases; Lead; Malignant Neoplasms; Measures; Mediating; Muscle Development; Mutation Analysis; Myotonia; Myotonic Dystrophy; Neuraxis; Oligonucleotides; Outcome; Pattern; Play; Proteins; RNA; RNA Binding; RNA Processing; RNA Sequences; RNA Splicing; RNA-Binding Proteins; Regulation; Reporter; Role; Site; Skeletal Development; Structure; Symptoms; System; Testing; Tissues; associated symptom; biomarker selection; cardiogenesis; combinatorial; human disease; in vivo; mRNA Precursor; nervous system disorder; next generation sequencing; novel; paralogous gene; predictive modeling; response; transcriptome

Many trans and cis acting factors that control alternative splicing have been identified. The explosion of next-generation sequencing approaches have identified thousands of regulated splicing events (RNA-seq), and binding sites of many proteins which regulate alternative splicing have been identified via CLIPseq. An important question remaining for the field is: What are the rules governing the behavior of alternative splicing decisions? For example, what are the RNA elements (sequence and structure) that determine if alternatively regulated exons will respond at low concentrations or at high concentrations to a splicing factor, and will the splicing responses exhibit cooperative behavior or not? Addressing these questions is important for providing a framework for understanding how changes in splicing factor concentration can lead to disease. To address these questions, we have created cellular models that allow us to precisely titrate the level of an alternative splicing regulator, the Muscleblind-like 1 (MBNL1) protein. MBNL1 has been shown to regulate thousands of alternative splicing events and is important for the development of skeletal muscle, heart and the central nervous system. This regulation is highlighted by the primary role that MBNL proteins play in the disease myotonic dystrophy (DM), in which MBNL1 and its paralogs (MBNL2 and MBNL3) are sequestered by expanded CUG or CCUG repeat RNAs, resulting in aberrant RNA processing. The mis- splicing of MBNL targets has been shown to be responsible for causing some of the symptoms associated with DM, including the hallmark symptom myotonia. .

控制替代剪接的许多跨和顺式作用因素具有 已确定。下一代测序方法的爆炸有 确定了成千上万的调节剪接事件（RNA-SEQ）和结合位点 许多调节替代剪接的蛋白质已通过 Clipseq。该领域剩下的一个重要问题是：规则是什么 管理替代剪接决策的行为？例如，什么是 RNA元素（序列和结构）确定是否替代 受调节的外显子将以低浓度或高浓度的响应 拼接因子，剪接响应将表现出合作行为或 不是？解决这些问题对于提供一个框架很重要 了解剪接因子浓度的变化如何导致疾病。 为了解决这些问题，我们创建了蜂窝模型，使我们能够 精确地滴定替代剪接调节器的水平，即肌肉闪烁 1（MBNL1）蛋白质。已显示MBNL1调节数千种替代方案 拼接事件，对于骨骼肌的发展很重要，心脏 和中枢神经系统。该法规由主要角色强调 MBNL蛋白在疾病中发挥的肌发育症（DM），其中 MBNL1及其旁系同源物（MBNL2和MBNL3）被扩展 CUG或CCUG重复RNA，导致异常RNA处理。误会 MBNL目标的拼接已被证明是造成某些的原因 与DM相关的症状，包括标志性症状肌瘤。 。