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Abstract：
Gene therapeutic approaches to cure genetic diseases require tools to express the rescuing gene exclusively within the affected tissues. Viruses are often chosen as gene transfer vehicles but they have limited capacity for genetic information to be carried and transduced. In addition, to avoid off-target effects the therapeutic gene should be driven by a tissue-specific promoter in order to ensure expression in the target organs, tissues, or cell populations. The larger the promoter, the less space will be left for the respective gene. Thus, there is a need for small but tissue-specific promoters. Here, we describe a compact unc45b promoter fragment of 195 bp that retains the ability to drive gene expression exclusively in skeletal and cardiac muscle in zebrafish and mouse. Remarkably, the described unc45b promoter fragment not only drives muscle-specific expression but presents heat-shock inducibility, allowing a temporal and spatial quantity control of (trans)gene expression. Here, we demonstrate that the transgenic expression of the smyd1b gene driven by the unc45b promoter fragment is able to rescue the embryonically lethal heart and skeletal muscle defects in smyd1b-deficient flatline mutant zebrafish. Our findings demonstrate that the described muscle-specific unc45b promoter fragment might be a valuable tool for the development of genetic therapies in patients suffering from myopathies. genesis 54:431-438, 2016. © 2016 The Authors. Genesis Published by Wiley Periodicals, Inc.
摘要：
治疗遗传疾病的基因治疗方法需要工具来仅在受影响的组织内表达拯救基因。通常选择病毒作为基因转移载体,但它们携带和转导遗传信息的能力有限。此外,为了避免脱靶效应，治疗基因应该由组织特异性启动子驱动，以确保在靶器官中表达，组织，或细胞群。发起人越大，为各自的基因留下的空间就越少。因此，需要小的但组织特异性的启动子。这里,我们描述了195bp的紧密的unc45b启动子片段，它保留了驱动基因表达的能力，只在斑马鱼和小鼠的骨骼肌和心肌中。值得注意的是,所描述的unc45b启动子片段不仅驱动肌肉特异性表达，而且呈现热休克诱导性，允许（反式）基因表达的时间和空间数量控制。这里,我们证明，由unc45b启动子片段驱动的smyd1b基因的转基因表达能够挽救smyd1b缺陷的扁平线突变体斑马鱼的胚胎致死性心脏和骨骼肌缺陷。我们的发现表明，所描述的肌肉特异性unc45b启动子片段可能是开发患有肌病的患者的遗传疗法的有价值的工具。创世纪54:431-438,2016.©2016作者创世纪由Wiley期刊出版，Inc.
