New York University Abu Dhabi, PO Box 129188, Abu Dhabi, UAE.
Department of Chemistry, University of Georgia, Athens, GA 30602, USA.
Molecules. 2020 Apr 29;25(9):2078. doi: 10.3390/molecules25092078.
Photoactivatable cyclic caged morpholino oligomers (ccMOs) represent a promising tool to selectively regulate gene expression with spatiotemporal control. Nevertheless, some challenges associated with the preparation of these reagents have limited their broader use in biological settings. We describe a novel ccMO design that overcomes many of the challenges and considerably expedites the synthetic preparation. The key factor is the introduction of an ethynyl function on the photocleavable linker to facilitate the use of a Huisgen 1,3-dipolar cycloaddition for the coupling reaction with the oligonucleotide. Compared to previous strategies, this modification reduces the number of synthetic steps and significantly improves the total yield and the stability of the linker. We used the alkynyl-functionalized linker for the preparation of two different ccMOs targeting the mRNA of the glutamic acid decarboxylase genes, and . HPLC analysis confirms that the caging strategy successfully inhibits the DNA binding ability, and the activity can be restored by brief illumination with 405-nm light. Overall, the straightforward preparation together with the clean and fast photochemistry make these caged antisense reagents excellent tools to modulate gene function in-vivo with spatial and temporal precision.
光可激活的环状笼状修饰型寡核苷酸（ccMOs）是一种很有前途的工具，可用于具有时空控制的选择性基因表达调控。然而，由于这些试剂的制备存在一些挑战，限制了它们在生物学环境中的更广泛应用。我们描述了一种新型 ccMO 设计，克服了许多挑战，并大大加快了合成准备过程。关键因素是在光可裂解连接子上引入乙炔基官能团，以促进与寡核苷酸的 Huisgen 1,3-偶极环加成反应的偶联反应。与以前的策略相比，这种修饰减少了合成步骤的数量，并显著提高了连接子的总产率和稳定性。我们使用炔基官能化的连接子来制备两种针对谷氨酸脱羧酶基因 mRNA 的不同 ccMO，和。HPLC 分析证实，笼状策略成功抑制了 DNA 结合能力，并且可以通过用 405nm 光短暂照射来恢复其活性。总的来说，这种简单的制备方法以及干净和快速的光化学反应使这些笼状反义试剂成为在体内具有空间和时间精度调节基因功能的优秀工具。
