Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA 15213, USA.
Development. 2023 Jan 1;150(1). doi: 10.1242/dev.201373. Epub 2023 Jan 6.
Sea urchins and other echinoderms are important experimental models for studying developmental processes. The lack of approaches for conditional gene perturbation, however, has made it challenging to investigate the late developmental functions of genes that have essential roles during early embryogenesis and genes that have diverse functions in multiple tissues. The doxycycline-controlled Tet-On system is a widely used molecular tool for temporally and spatially regulated transgene expression. Here, we optimized the Tet-On system to conditionally induce gene expression in sea urchin embryos. Using this approach, we explored the roles the MAPK signaling plays in skeletogenesis by expressing genes that perturb the pathway specifically in primary mesenchyme cells during later stages of development. We demonstrated the wide utility of the Tet-On system by applying it to a second sea urchin species and in cell types other than the primary mesenchyme cells. Our work provides a robust and flexible platform for the spatiotemporal regulation of gene expression in sea urchins, which will considerably enhance the utility of this prominent model system.
海胆和其他棘皮动物是研究发育过程的重要实验模型。然而，缺乏条件基因干扰的方法，使得研究在早期胚胎发生中具有重要作用的基因和在多种组织中具有多种功能的基因的晚期发育功能具有挑战性。强力霉素控制的 Tet-On 系统是一种广泛用于时空调节转基因表达的分子工具。在这里，我们优化了 Tet-On 系统，以在海胆胚胎中条件性诱导基因表达。使用这种方法，我们通过在发育后期的初级间充质细胞中特异性表达干扰该途径的基因，探索了 MAPK 信号在骨骼发生中的作用。我们通过将其应用于第二种海胆物种和初级间充质细胞以外的细胞类型，证明了 Tet-On 系统的广泛适用性。我们的工作为海胆中的基因表达提供了一个强大而灵活的时空调节平台，这将极大地增强这个重要模型系统的实用性。
