PDF(Pubmed)
Abstract：
Rod photoreceptor formation in the postnatal mouse is a widely used model system for studying mammalian photoreceptor development. This experimental paradigm provides opportunities for both gain and loss-of-function studies which can be accomplished through in vivo plasmid delivery and electroporation. However, the cis-regulatory elements used to implement this approach have not been fully evaluated or optimized for the unique transcriptional environment of photoreceptors. Here we report that the use of a photoreceptor cis-regulatory element from the Crx gene in combination with broadly active promoter elements can increase the targeting of developing rod photoreceptors in the mouse. This can lead to greater reporter expression, as well as enhanced misexpression and loss-of-function phenotypes in these cells. This study also highlights the importance of identifying and testing relevant cis-regulatory elements when planning cell subtype specific experiments. The use of the specific hybrid elements in this study will provide a more efficacious gene delivery system to study mammalian photoreceptor formation.
摘要：
出生后小鼠中的杆感光体形成是用于研究哺乳动物感光体发育的广泛使用的模型系统。该实验范式为功能获得和功能丧失研究提供了机会，这些研究可以通过体内质粒递送和电穿孔来完成。然而,用于实施该方法的顺式调控元件尚未针对光感受器的独特转录环境进行充分评估或优化.在这里，我们报道了来自Crx基因的光感受器顺式调节元件与广泛活性的启动子元件结合使用可以增加小鼠发育中的杆状光感受器的靶向性。这可以导致更大的记者表达，以及这些细胞中增强的错误表达和功能丧失表型。这项研究还强调了在计划细胞亚型特异性实验时识别和测试相关顺式调控元件的重要性。在这项研究中使用特定的杂合元件将为研究哺乳动物感光体形成提供更有效的基因递送系统。
