Abstract
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.
Highlights
Notch signaling is highly conserved in most animals and plays critical roles during neurogenesis as well as embryonic development
After scFv recognizes the antigen on the sender cells, a conformational change of the negative regulatory region (NRR) in Notch core relays the signal to the Notch transmembrane domain (TMD) in Notch core
We further showed that adding an intracellular hydrophobic sequence (QHGQLWF) after Notch core significantly reduces ligand-independent activation (LIA) of synNotch, without affecting the efficiency of antigen-induced activation efficiency
Summary
Notch signaling is highly conserved in most animals and plays critical roles during neurogenesis as well as embryonic development. Customizing cells to sense an extracellular signal and respond is an important objective in synthetic biology To this end, a variety of tools capable of transmembrane signaling have been developed, such as the chimeric antigen receptor[1], synthetic Notch[2,3,4], MESA5, Tango[6], ChaCha[7], and GEMS8. We further showed that adding an intracellular hydrophobic sequence (QHGQLWF) after Notch core significantly reduces LIA of synNotch, without affecting the efficiency of antigen-induced activation efficiency. We confirmed this improvement with multiple variants of synNotch, and named our improved version the enhanced synthetic Notch receptor, esNotch
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
