Department of Biochemistry, School of Life Sciences and Zhongshan Hospital, Fudan University, 200438, Shanghai, China.
School of Basic Medical Sciences, Fudan University, Shanghai, China.
Commun Biol. 2020 Mar 13;3(1):116. doi: 10.1038/s42003-020-0848-x.
Notch signaling is highly conserved in most animals and plays critical roles during neurogenesis as well as embryonic development. Synthetic Notch-based systems, modeled from Notch receptors, have been developed to sense and respond to a specific extracellular signal. Recent advancement of synNotch has shown promise for future use in cellular engineering to treat cancers. However, synNotch from Morsut et al. (2016) has a high level of ligand-independent activation, which limits its application. Here we show that adding an intracellular hydrophobic sequence (QHGQLWF, named as RAM7) present in native Notch, significantly reduced ligand-independent activation. Our enhanced synthetic Notch receptor (esNotch) demonstrates up to a 14.6-fold reduction in ligand-independent activation, without affecting its antigen-induced activation efficiency. Our work improves a previously reported transmembrane receptor and provides a powerful tool to develop better transmembrane signaling transduction modules for further advancement of eukaryotic synthetic biology.
Notch 信号通路在大多数动物中高度保守，在神经发生和胚胎发育过程中发挥着关键作用。基于 Notch 受体设计的合成 Notch 系统已被开发出来，用于感知和响应特定的细胞外信号。最近，synNotch 的进展为未来在细胞工程中治疗癌症提供了希望。然而，Morsut 等人（2016 年）的 synNotch 具有高水平的配体非依赖性激活，限制了其应用。在这里，我们表明添加存在于天然 Notch 中的细胞内疏水序列（QHGQLWF，命名为 RAM7）可显著降低配体非依赖性激活。我们的增强型合成 Notch 受体（esNotch）显示配体非依赖性激活降低了 14.6 倍，而不影响其抗原诱导的激活效率。我们的工作改进了之前报道的跨膜受体，并提供了一个强大的工具来开发更好的跨膜信号转导模块，以进一步推进真核合成生物学。
