PLP Splicing: In Vivo Analysis in A Mouse Model

描述（由申请人提供）：本项目的长期目标是阐明PLP基因内含子3中富含G的19 bp序列在体内PLP-DM 20剪接位点选择中的功能。 研究人员已经表明，该序列的缺失会导致人类神经系统疾病，并且该序列在体外调节少突胶质细胞中PLP剪接的选择。 富含G的元件已被证明在体外调节嵌合基因中的剪接位点选择。 研究人员建议产生一系列携带富含G的19 bp序列缺失的敲入小鼠，并利用这些小鼠研究发育过程中大脑和外周神经中PLP的特异性剪接。 在这些研究中，研究人员将在髓鞘形成的早期阶段、髓鞘形成的高峰期和成年期描述剪接的特征。 将研究PLP和其他髓鞘蛋白的产生以及髓鞘形成。 小鼠表型将通过运动试验表征。 据研究者所知，设计用于研究可变剪接的体内调节的小鼠模型不可用，并且没有PLP小鼠突变体允许研究PLP剪接。 待制备的小鼠模型将允许在整个基因的背景下和在存在外部线索的情况下研究PLP基因中的选择性剪接的调节。 此外，它有可能产生关于许多其他基因剪接的更广泛的生物学相关性数据。

DESCRIPTION (provided by applicant): The long-term goals of this project are to elucidate the function of a G-rich 19-bp sequence in intron 3 of the PLP gene in PLP-DM20 splice site selection in vivo. The investigators have shown that deletion of this sequence causes a neurological disease in humans and that this sequence regulates selection of PLP splicing in oligodendrocytes in vitro. G-rich elements have been shown to regulate splice site selection in chimeric genes in vitro. The investigators propose to generate a line of knock-in mice carrying deletion of the G-rich 19-bp sequence and to utilize these mice to investigate PLP specific splicing in brain and peripheral nerves during development. In these studies, the investigators will characterize splicing at the early stages of myelination, peak of myelination, and adulthood. The production of PLP and other myelin proteins and myelin formation will be studied. The mouse phenotype will be characterized by motor testing. To the investigators' knowledge, mouse models designed to investigate in vivo regulation of alternative splicing are not available and none of the PLP mouse mutants allows studies of PLP splicing. The mouse model to be made will allow investigating the regulation of alternative splicing in the PLP gene in the context of the entire gene and in the presence of external cues. In addition, it has the potential to generate data of broader biological relevance about splicing in many other genes.