Weissenboeck Florian P, Pieper Melissa, Schepers Helena, Hötte Sophie, Klöcker Nils, Hüwel Sabine, van Impel Andreas, Schulte-Merker Stefan, Rentmeister Andrea
Institute of Biochemistry, University of Münster, Münster, Germany.
Institute for Cardiovascular Organogenesis and Regeneration, Faculty of Medicine, University of Münster, Münster, Germany.
Commun Chem. 2025 Jan 19;8(1):16. doi: 10.1038/s42004-025-01411-7.
Translation of mRNA into protein is a fundamental process and tightly controlled during development. Several mechanisms acting on the mRNA level regulate when and where an mRNA is expressed. To explore the effects of conditional and transient gene expression in a developing organism, it is vital to experimentally enable abrogation and restoration of translation. We recently developed the FlashCaps technology allowing preparation of translationally muted mRNAs and their controlled activation by light. Here, we validate its functionality in vivo. We demonstrate that translation of FlashCap-eGFP-mRNA can be triggered in zebrafish embryos with spatiotemporal control. The injected FlashCap-mRNA is stable for hours and remains muted. Light-mediated activation up to 24 h post fertilization produces visible amounts of eGFP and can be restricted to distinct parts of the embryo. This methodology extends the toolbox for vertebrate models by enabling researchers to locally activate mRNA translation at different timepoints during development.
信使核糖核酸（mRNA）翻译成蛋白质是一个基本过程，在发育过程中受到严格控制。几种作用于mRNA水平的机制调节着mRNA表达的时间和位置。为了探究发育中的生物体中条件性和瞬时性基因表达的影响，通过实验实现翻译的废除和恢复至关重要。我们最近开发了FlashCaps技术，可制备翻译沉默的mRNA并通过光对其进行可控激活。在此，我们在体内验证了其功能。我们证明，FlashCap-eGFP-mRNA的翻译可在斑马鱼胚胎中通过时空控制触发。注射的FlashCap-mRNA可稳定数小时并保持沉默状态。受精后长达24小时的光介导激活可产生可见量的绿色荧光蛋白（eGFP），并且可以局限于胚胎的不同部位。这种方法通过使研究人员能够在发育过程的不同时间点局部激活mRNA翻译，扩展了脊椎动物模型的工具箱。
