Department of Chemistry, University of Pittsburgh, Pittsburgh, PA, USA.
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA, USA.
Dev Biol. 2021 Jul;475:21-29. doi: 10.1016/j.ydbio.2021.02.014. Epub 2021 Mar 5.
Echinoderms are important experimental models for analyzing embryonic development, but a lack of spatial and temporal control over gene perturbations has hindered developmental studies using these animals. Morpholino antisense oligonucleotides (MOs) have been used successfully by the echinoderm research community for almost two decades, and MOs remain the most widely used tool for acute gene knockdowns in these organisms. Echinoderm embryos develop externally and are optically transparent, making them ideally-suited to many light-based approaches for analyzing and manipulating development. Studies using zebrafish embryos have demonstrated the effectiveness of photoactivatable (caged) MOs for conditional gene knockdowns. Here we show that caged MOs, synthesized using nucleobase-caged monomers, provide light-regulated control over gene expression in sea urchin embryos. Our work provides the first robust approach for conditional gene silencing in this prominent model system.
棘皮动物是分析胚胎发育的重要实验模型，但由于缺乏对基因干扰的时空控制，这些动物的发育研究受到了阻碍。近二十年来，反义寡核苷酸（MOs）在棘皮动物研究领域得到了成功应用，并且 MOs 仍然是这些生物中急性基因敲低最广泛使用的工具。棘皮动物的胚胎是体外发育的，并且是光学透明的，这使得它们非常适合使用许多基于光的方法来分析和操作发育。使用斑马鱼胚胎的研究表明，光激活（笼状）MOs 可用于条件性基因敲低。在这里，我们展示了使用碱基笼状单体合成的笼状 MOs 可提供对海胆胚胎中基因表达的光调控控制。我们的工作为这个重要的模式系统提供了第一个稳健的条件性基因沉默方法。
