Abstract
Cell-type diversity in multicellular organisms is created through a series of binary cell fate decisions. Lateral inhibition controlled by Delta-Notch signalling is the core mechanism for the choice of alternative cell types by homogeneous neighbouring cells. Here, we show that cells engineered with a Delta-Notch-dependent lateral inhibition circuit spontaneously bifurcate into Delta-positive and Notch-active cell populations. The synthetic lateral inhibition circuit comprises transcriptional repression of Delta and intracellular feedback of Lunatic fringe (Lfng). The Lfng-feedback subcircuit, even alone, causes the autonomous cell-type bifurcation. Furthermore, the ratio of two cell populations bifurcated by lateral inhibition is reproducible and robust against perturbation. The cell-type ratio is adjustable by the architecture of the lateral inhibition circuit as well as the degree of cell-cell attachment. Thus, the minimum lateral inhibition mechanism between adjacent cells not only serves as a binary cell-type switch of individual cells but also governs the cell-type ratio at the cell-population level.
Highlights
Cell-type diversity in multicellular organisms is created through a series of binary cell fate decisions
All of the genetic circuits were constructed in Chinese hamster ovary (CHO) cells, which do not have a native lateral inhibition mechanism
Using the aTRLF cells, we investigated whether the ratio of red cells to green cells is reproducible, or robust, as robustness of cell-type ratios should be pivotal for the development of multicellular organisms
Summary
Cell-type diversity in multicellular organisms is created through a series of binary cell fate decisions. We show that cells engineered with a Delta–Notch-dependent lateral inhibition circuit spontaneously bifurcate into Delta-positive and Notch-active cell populations. Considering the fact that more than 40 arbitrary fluorescence units (AFUs) of the tTS-2A-GFP signal are required to effectively repress the PTetO activity (Supplementary Fig. 1d), we selected cell clones where more than 100 AFU of tTS-2A-GFP were induced from the PTP1 (Supplementary Fig. 1e). To this end, the PTP1-tTS-2A-GFP construct was introduced by an efficient transposon vector, piggyBac[26], while other constructs were introduced by a lentiviral vector. As a result of these improvements, a PTetO-luciferase reporter construct showed an 18-fold decrease on activation of Notch (Supplementary Fig. 3a)
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
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.
