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: 9902459
• 负责人: Andrew Berglund
• 金额: $ 41.44万
• 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 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9902459
• 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; Heart Diseases; Lead; Malignant Neoplasms; Measures; Mediating; 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; Skeletal Muscle; Structure; Symptoms; System; Testing; Tissues; associated symptom; biomarker selection; combinatorial; human disease; in vivo; mRNA Precursor; nervous system disorder; next generation sequencing; novel; paralogous gene; predictive modeling; response; transcriptome; transcriptome sequencing

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. .

许多控制选择性剪接的反式和顺式作用因子 已被识别。新一代测序方法的爆炸式增长 鉴定了数千个受调控的剪接事件（RNA-seq）以及 许多调节选择性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 控制选择性剪接决策的行为？例如，什么是 决定是否可选择的 RNA 元件（序列和结构） 受调控的外显子会在低浓度或高浓度下对 剪接因子，以及剪接反应是否表现出合作行为或 不是？解决这些问题对于提供一个框架非常重要 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，我们创建了细胞模型，使我们能够 精确滴定另一种剪接调节因子（Muscleblind-like）的水平 1 (MBNL1) 蛋白。 MBNL1 已被证明可以调节数千种替代品 剪接事件，对于骨骼肌、心脏的发育很重要 和中枢神经系统。该法规的主要作用是强调 MBNL 蛋白在强直性肌营养不良 (DM) 疾病中发挥作用，其中 MBNL1 及其旁系同源物（MBNL2 和 MBNL3）通过扩展被隔离 CUG 或 CCUG 重复 RNA，导致 RNA 加工异常。错误的是 MBNL 靶标的剪接已被证明是导致某些 与 DM 相关的症状，包括标志性症状肌强直。 。