Istituto Italiano di Tecnologia-IIT, Largo Barsanti e Matteucci, 80125, Naples, Italy.
University of Genoa, 16132, Genoa, Italy.
Nat Commun. 2018 Oct 22;9(1):4392. doi: 10.1038/s41467-018-06825-7.
The development of RNA-encoded regulatory circuits relying on RNA-binding proteins (RBPs) has enhanced the applicability and prospects of post-transcriptional synthetic network for reprogramming cellular functions. However, the construction of RNA-encoded multilayer networks is still limited by the availability of composable and orthogonal regulatory devices. Here, we report on control of mRNA translation with newly engineered RBPs regulated by viral proteases in mammalian cells. By combining post-transcriptional and post-translational control, we expand the operational landscape of RNA-encoded genetic circuits with a set of regulatory devices including: i) RBP-protease, ii) protease-RBP, iii) protease-protease, iv) protein sensor protease-RBP, and v) miRNA-protease/RBP interactions. The rational design of protease-regulated proteins provides a diverse toolbox for synthetic circuit regulation that enhances multi-input information processing-actuation of cellular responses. Our approach enables design of artificial circuits that can reprogram cellular function with potential benefits as research tools and for future in vivo therapeutics and biotechnological applications.
基于 RNA 结合蛋白（RBPs）的 RNA 编码调控回路的发展增强了转录后合成网络在重新编程细胞功能方面的适用性和前景。然而，RNA 编码多层网络的构建仍然受到可组合和正交调控装置的可用性的限制。在这里，我们报告了在哺乳动物细胞中利用新工程化的受病毒蛋白酶调控的 RBPs 来控制 mRNA 翻译的情况。通过结合转录后和翻译后调控，我们用一组调控装置扩展了 RNA 编码遗传回路的操作范围，这些调控装置包括：i）RBP-蛋白酶，ii）蛋白酶-RBP，iii）蛋白酶-蛋白酶，iv）蛋白传感器-蛋白酶-RBP，以及 v）miRNA-蛋白酶/RBP 相互作用。受蛋白酶调控的蛋白质的合理设计为合成回路调控提供了多样化的工具包，增强了细胞反应的多输入信息处理-触发。我们的方法能够设计出具有人工电路，可以重新编程细胞功能，作为研究工具，以及未来体内治疗和生物技术应用具有潜在的好处。
