(A) Abstract diagram of MINPA topology, where X and Y mutually inhibit (T-bars) each other and auto-activate (arrowheads) itself. Four inducers to regulate the four color-coded regulatory edges are …
(A) Abstract diagrams and molecular implementation of the eight MINPA sub-network topologies, including tunable positive feedbacks (T6 and T9), mutual inhibition (T5), dual-positive feedbacks (T10), …
(A) Schematic illustration of rationale for sequential induction. This two-parameter bifurcation diagram of a bistable toggle-switch depicts all steady state values of response (Z-axis) with …
(A) Abstract diagram and molecular implementation of the toggle switch circuit. TetR (R) and LacI (I) mutually inhibit each other through binding to Ptet and Plac promoter, respectively. IPTG and …
(A) Arabinose (Ara) and IPTG were sequentially (left and middle columns) or simultaneously (right column) applied to induce T15. The first inducer was applied for 5 hr, and then the second inducer …
Left: IPTG was first applied to induce the circuits, and then aTc was added; Middle: aTc was first applied to induce the circuits, and then IPTG was added; Right: IPTG and aTc were added …
(A) Engineered MINPA is tunable to reach four individual states: low-low, GFP, mCherry, and high-high, under no induction, 1*10−4 M AHL and 100 ng/ml aTc, 2.5*10−5 (m/v) Arabinose, 1*10−4 M AHL and …
The circuit’s quadrastability is illustrated as four similar-sized colored spheres on the same gray plane, which represents the low-low, GFP, mCherry, and high-high state, respectively. Blue lines …
(A–D) One-dimensional bifurcation analysis for all four inducers in MINPA. We perform bifurcation analysis for each inducer while setting the concentration of other inducers to be very small (10−10).…
(A) Dynamic evolution of computed energy landscapes of MINPA under sequential/simultaneous inductions of Arabinose, and/or AHL and aTc. Center route: simultaneous induction with three inducers; Left …
(A) Up left: Flow cytometry result for cells simultaneously induced with 2.5*10−5m/v Arabinose, 1*10−4 M AHL and 200 ng/ml aTc for 6.5 hr. Up right: Cells were first induced with 1*10−4 M and 400 …
Growth curves for the cells under the four individual inducers: Ara; AHL; aTc; IPTG, and inducer combinations: AHL and aTc; AHL, aTc, and Ara. Ara: 2.5*10−5m/v; AHL: 1*10−5 M, aTc: 200 ng/ml, IPTG: …
Time course of the cells growing and fluorescence state change with 2*10−4 M IPTG and 200 ng/ml aTc induction in the trap. The red flow is medium without inducer for 6 hr, and then cells switch to …
Components from the Registry of standard biological parts
Biobrick number | Abbreviation in the paper | Description |
---|---|---|
BBa_C0080 | AraC | AraC arabinose operon regulatory protein from E. coli |
BBa_C0040 | TetR | Tetracycline repressor from transposon Tn10 |
BBa_C0062 | LuxR | LuxR activator from Aliivibrio fischeri |
BBa_C0012 | LacI | LacI repressor from E. coli |
BBa_E0240 | GFP | GFP generator |
BBa_J06702 | mCherry | RFP generator |
BBa_K176002 | Plux/tet | Hybrid promoter with LuxR/HSL- and TetR-binding sites |
BBa_B0034 | RBS | Ribosome binding site |
BBa_B0015 | Terminator | Transcriptional terminator (double) |
BBa_K176009 | CP | Constitutive promoter |
pSB1K3 | pSB1K3 | High copy BioBrick assembly plasmid with kanamycin resistance |
pSB1A3 | pSB1A3 | High copy BioBrick assembly plasmid with ampicillin resistance |
Flow cytometry data analysis file.
Bifurcation analysis in Figure 3.
Potential landscape calculation file.