Modulation of alternative splicing of trafficking genes by genome editing reveals functional consequences in muscle biology
Modulation of alternative splicing of trafficking genes by genome editing reveals functional consequences in muscle biology
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DOI：
10.1016/j.biocel.2018.10.004
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发表时间：
2018-12-01
影响因子：
4
通讯作者：
Giudice, Jimena
中科院分区：
文献类型：
--
作者：
Blue, R. Eric;Koushik, Amrita;Giudice, Jimena
选择性剪接是一种调控机制，通过它从单个基因产生多种mRNA亚型。许多编码膜运输蛋白的基因是选择性剪接的。然而，有有限的信息，从这些剪接过渡导致的功能后果。在这里，我们开发了适当的工具来研究选择性剪接在大多数体内环境中对发育的功能影响。其次，我们提供了肌肉发育过程中剪接调控的生理意义的证据。我们以前在小鼠心脏发育中的工作确定了三个由出生和成年之间的选择性剪接调控的贩运基因：网格蛋白重链，网格蛋白轻链-a和贩运驱动蛋白结合蛋白-1。在这里，我们证明了这三个基因的选择性剪接调控是组织和发育阶段特异性的。为了确定体内剪接调控的功能后果，我们使用基因组编辑来阻断胚胎至成体的剪接转换。我们表征了这些小鼠品系之一的表型，并证明了当网格蛋白重链基因的剪接调节被阻止时，小鼠表现出体重和肌肉重量的增加，这是由于肌纤维尺寸的增大。这项工作的意义有两个方面。首先，我们揭示了网格蛋白重链在肌肉生长中的新作用，并表明其通过选择性剪接的调节有助于肌肉发育。第二，新的小鼠品系将提供一个有用的工具，研究如何剪接调节三个运输基因影响组织身份的收购和成熟在体内。
Alternative splicing is a regulatory mechanism by which multiple mRNA isoforms are generated from single genes. Numerous genes that encode membrane trafficking proteins are alternatively spliced. However, there is limited information about the functional consequences that result from these splicing transitions. Here, we developed appropriate tools to study the functional impact of alternative splicing in development within the most in vivo context. Secondly, we provided evidence of the physiological implications of splicing regulation during muscle development. Our previous work in mouse heart development identified three trafficking genes that are regulated by alternative splicing between birth and adulthood: the clathrin heavy chain, the clathrin light chain-a, and the trafficking kinesin binding protein-1. Here, we demonstrated that alternative splicing regulation of these three genes is tissue- and developmental stage-specific. To identify the functional consequences of splicing regulation in vivo, we used genome editing to block the neonatal-to-adult splicing transitions. We characterized the phenotype of one of these mouse lines and demonstrated that when splicing regulation of the clathrin heavy chain gene is prevented mice exhibit an increase in body and muscle weights which is due to an enlargement in myofiber size. The significance of this work has two components. First, we revealed novel roles of the clathrin heavy chain in muscle growth and showed that its regulation by alternative splicing contributes to muscle development. Second, the new mouse lines will provide a useful tool to study how splicing regulation of three trafficking genes affects tissue identity acquisition and maturation in vivo.