Biocenter, Johannes Gutenberg University, Mainz, Germany.
Departments of Biology and Chemistry, IMPRS on Cellular Biophysics, Mainz, Germany.
Nat Commun. 2024 Oct 24;15(1):9177. doi: 10.1038/s41467-024-53403-1.
RNA modifications play a pivotal role in the regulation of RNA chemistry within cells. Several technologies have been developed with the goal of using RNA modifications to regulate cellular biochemistry selectively, but achieving selective and precise modifications remains a challenge. Here, we show that by using designer organelles, we can modify mRNA with pseudouridine in a highly selective and guide-RNA-dependent manner. We use designer organelles inspired by concepts of phase separation, a central tenet in developing artificial membraneless organelles in living mammalian cells. In addition, we use circular guide RNAs to markedly enhance the effectiveness of targeted pseudouridinylation. Our studies introduce spatial engineering through optimized RNA editing organelles (OREO) as a complementary tool for targeted RNA modification, providing new avenues to enhance RNA modification specificity.
RNA 修饰在调节细胞内的 RNA 化学中起着关键作用。已经开发了几种技术，旨在利用 RNA 修饰来有选择地调节细胞生物化学，但实现选择性和精确的修饰仍然是一个挑战。在这里，我们表明，通过使用设计的细胞器，我们可以以高度选择性和依赖向导 RNA 的方式用假尿嘧啶核苷修饰 mRNA。我们使用受相分离概念启发的设计细胞器，这是在活哺乳动物细胞中开发无膜细胞器的核心原则。此外，我们还使用环形向导 RNA 显著提高了靶向假尿嘧啶核苷修饰的效果。我们的研究通过优化 RNA 编辑细胞器（OREO）引入空间工程，作为靶向 RNA 修饰的补充工具，为增强 RNA 修饰特异性提供了新途径。
