Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, United States.
Department of Developmental Biology, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, United States.
J Am Chem Soc. 2021 Nov 10;143(44):18665-18671. doi: 10.1021/jacs.1c08723. Epub 2021 Oct 27.
Conditionally activated, caged morpholino antisense agents (cMOs) are tools that enable the temporal and spatial investigation of gene expression, regulation, and function during embryonic development. Cyclic MOs are conformationally gated oligonucleotide analogs that do not block gene expression until they are linearized through the application of an external trigger, such as light or enzyme activity. Here, we describe the first examples of small molecule-responsive cMOs, which undergo rapid and efficient decaging via a Staudinger reduction. This is enabled by a highly flexible linker design that offers opportunities for the installation of chemically activated, self-immolative motifs. We synthesized cyclic cMOs against two distinct, developmentally relevant genes and demonstrated phosphine-triggered knockdown of gene expression in zebrafish embryos. This represents the first report of a small molecule-triggered antisense agent for gene knockdown, adding another bioorthogonal entry to the growing arsenal of gene knockdown tools.
条件激活的、笼状的吗啉代反义寡核苷酸（cMO）是一种工具，可在胚胎发育过程中实现对基因表达、调控和功能的时空研究。环状 MO 是构象门控的寡核苷酸类似物，只有在通过外部触发（如光或酶活性）线性化后，才会阻断基因表达。在这里，我们描述了第一个小分子响应的 cMO 的例子，它们通过施蒂德格还原反应快速有效地去笼。这得益于高度灵活的连接子设计，为安装化学激活的自毁性基团提供了机会。我们合成了针对两个不同的、发育相关基因的环状 cMO，并在斑马鱼胚胎中证明了磷烷触发的基因表达下调。这代表了第一个小分子触发的反义药物用于基因敲低的报告，为不断增长的基因敲低工具库增加了另一种生物正交的方法。
