Transcriptome processing networks in skeletal muscle: mechanisms and functions

骨骼肌转录组处理网络：机制和功能

• 批准号: 10585923
• 负责人: Thomas A Cooper
• 金额: $ 45.06万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10585923
• 关键词: Actin-Binding Protein; Actins; Adult; Affect; Age; Alleles; Alternative Splicing; Amino Acids; Animals; Antibodies; Antisense Oligonucleotides; Architecture; Binding Proteins; CRISPR/Cas technology; Cells; Cellular Structures; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Data; Development; Disease; Electroporation; Environment; Epitopes; Event; Excision; Exons; Failure; Funding; Generations; Genes; Genomics; Goals; Homeostasis; Immunofluorescence Microscopy; Individual; Injury; Knowledge; Label; Length; MAP4; Mediating; Microscopic; Microscopy; Microtubules; Modification; Molecular Analysis; Monitor; Mus; Muscle; Muscle Development; Muscle Weakness; Muscle function; Neuromuscular Junction; Nuclear; Organelles; Pathogenesis; Physiological; Protein Dynamics; Protein Isoforms; Proteins; Publishing; RNA Splicing; Resolution; Role; Skeletal Muscle; Specificity; Stress; Striated Muscles; Structure; Tissues; Transport Process; Variant; Work; alpha Tubulin; c-myc Genes; calponin; density; fetal; flexor digitorum brevis; gene function; in vivo; insight; mRNA Precursor; postnatal; postnatal development; repaired; response; skeletal muscle differentiation; skeletal muscle weakness; transcriptome; zygote

Project Summary The long-term goal of this project is to determine the extent, regulatory mechanisms, and functional consequences of regulated pre-mRNA processing in skeletal muscle. A large number of genes express pre- mRNAs that undergo alternative splicing to produce conserved muscle-specific protein isoforms the functions of which are unknown. These isoforms often appear during late fetal or early postnatal development. Disruption of alternative splicing is a common feature of diseases affecting skeletal muscle, often involving reversion to fetal isoforms, yet little is known about the contributions of these changes to pathogenesis. An underlying hypothesis of this proposal is that a focus on genes with conserved fetal and adult protein isoforms, including adult isoforms that are muscle-specific, will discover not only previously unknown isoform-specific functions but also previously unknown gene functions critical for adult muscle homeostasis. To identify the functions of alternative splicing events in vivo we are using CRISPR-mediated removal of exons that undergo muscle- specific and/or postnatally regulated inclusion. In the first aim, we will determine the functions of a striated muscle-specific isoform of the Map4 microtubule-binding protein in which deletion of the muscle-specific exon significantly disrupts microtubule architecture in skeletal muscle myofibers and impacts muscle force generation. In the second aim we will determine the function of the skeletal muscle-specific Limch1 isoform, the absence of which also decreases skeletal muscle function in vivo. In the third aim we will use CRISPR- mediated introduction of epitope tags into endogenous genes to monitor the temporal and spatial details of the alternative splicing transitions and adult muscle-specific isoforms at the level of individual cells in vivo. This study is expected to identify previously unknown gene functions, increase understanding of adult skeletal muscle homeostasis and the impact of its disruption in disease.

项目概要 该项目的长期目标是确定范围、监管机制和功能 骨骼肌中受调节的前 mRNA 加工的后果。大量基因表达预 经过选择性剪接以产生保守的肌肉特异性蛋白质亚型的 mRNA 具有以下功能 其中未知。这些亚型通常出现在胎儿晚期或产后早期发育期间。破坏 选择性剪接是影响骨骼肌的疾病的一个共同特征，通常涉及逆转 胎儿亚型，但人们对这些变化对发病机制的贡献知之甚少。底层的 该提案的假设是重点关注具有保守胎儿和成人蛋白质亚型的基因，包括 肌肉特异性的成人亚型不仅会发现以前未知的亚型特异性功能，而且 还有以前未知的对成人肌肉稳态至关重要的基因功能。识别功能 体内的选择性剪接事件我们正在使用 CRISPR 介导的外显子去除，这些外显子经历了肌肉- 特定的和/或产后调节的包含。在第一个目标中，我们将确定条纹的功能 Map4 微管结合蛋白的肌肉特异性亚型，其中肌肉特异性外显子的缺失 显着破坏骨骼肌肌纤维的微管结构并影响肌肉力量 一代。在第二个目标中，我们将确定骨骼肌特异性 Limch1 亚型的功能， 缺乏它也会降低体内骨骼肌功能。在第三个目标中，我们将使用 CRISPR- 介导将表位标签引入内源基因以监测基因的时间和空间细节 体内单个细胞水平的选择性剪接转变和成人肌肉特异性亚型。这 研究预计将识别以前未知的基因功能，增加对成人骨骼的了解 肌肉稳态及其破坏对疾病的影响。

项目成果

Combinatorial effects of ion channel mis-splicing as a cause of myopathy in myotonic dystrophy.

• DOI: 10.1172/jci176089
• 发表时间: 2024-01-02
• 期刊: JOURNAL OF CLINICAL INVESTIGATION
• 影响因子: 15.9
• 作者: Nitschke, Larissa;Cooper, Thomas A.
• 通讯作者: Cooper, Thomas A.