Synthetic biology tools for engineering Goodwin oscillation in Trypanosoma brucei brucei.

Kinetoplastid protozoa possess properties that are highly divergent from the mammalian, yeast and bacterial cells more commonly used in synthetic biology and represent a tantalisingly untapped source of bioengineering potential. Trypanosoma brucei brucei (T. b. brucei), an established model organism for studying the Kinetoplastida, is non-pathogenic to humans and provides an interesting test case for establishing synthetic biology in this phylogenetic class. To demonstrate further the tractability of Kinetoplastida to synthetic biology, we sought to construct and demonstrate a Goodwin oscillator, the simplest oscillatory gene network, in T. b. brucei for the first time. We report one completed iteration of the archetypal synthetic biology Design-Build-Test-Learn (DBTL) cycle; firstly, using Ab initio mathematical modelling of the behaviour a theoretical, oscillatory, trypanosomal synthetic gene network (SGN) to inform the design of a plasmid encoding that network. Once assembled, the plasmid was then used to generate a stable transfectant T. b. brucei cell line. To test the performance of the oscillatory SGN, a novel experimental setup was established to capture images of the fluorescent signal from motion-restricted live cells. Data captured were consistent with oscillatory behaviour of the SGN, with cellular fluorescence observed to oscillate with a period of 50 min, with varying amplitude and linear growth trend. This first DBTL cycle establishes a foundation for future cycles in which the SGN design and experimental monitoring setup can be further refined.

Quantized cycling time in artificial gene networks induced by noise and intercell communication.

We propose a mechanism for the quantized cycling time based on the interplay of cell-to-cell communication and stochasticity, by investigating a model of coupled genetic oscillators with known topology. In addition, we discuss how inhomogeneity can be used to enhance such quantizing effects, while the degree of variability obtained can be controlled using the noise intensity or adequate system parameters.

Noise-memory induced excitability and pattern formation in oscillatory neural models.

We report a noise-memory induced phase transition in an array of oscillatory neural systems, which leads to the suppression of synchronous oscillations and restoration of excitable dynamics. This phenomenon is caused by the systematic contributions of temporally correlated parametric noise, i.e., possessing a memory, which stabilizes a deterministically unstable fixed point. Changing the noise correlation time, a reentrant phase transition to noise-induced excitability is observed in a globally coupled array. Since noise-induced excitability implies the restoration of the ability to transmit information, associated spatiotemporal patterns are observed afterwards. Furthermore, an analytic approach to predict the systematic effects of exponentially correlated noise is presented and its results are compared with the simulations.

Quantification of spatial structure of human proximal tibial bone biopsies using 3D measures of complexity.

Changes in trabecular bone composition during development of osteoporosis are used as a model for bone loss in microgravity conditions during a space flight. Symbolic dynamics and measures of complexity are proposed and applied to assess quantitatively the structural composition of bone tissue from 3D data sets of human tibia bone biopsies acquired by a micro-CT scanner. In order to justify the newly proposed approach, the measures of complexity of the bone architecture were compared with the results of traditional 2D bone histomorphometry. The proposed technique is able to quantify the structural loss of the bone tissue and may help to diagnose and to monitor changes in bone structure of patients on Earth as well as of the space-flying personnel.

Multistability and clustering in a population of synthetic genetic oscillators via phase-repulsive cell-to-cell communication.

We show that phase-repulsive coupling eliminates oscillations in a population of synthetic genetic clocks. For this, we propose an experimentally feasible synthetic genetic network that contains phase repulsively coupled repressilators with broken temporal symmetry. As the coupling strength increases, silencing of oscillations is found to occur via the appearance of an inhomogeneous limit cycle, followed by oscillation death. Two types of oscillation death are observed: For lower couplings, the cells cluster in one of two stationary states of protein expression; for larger couplings, all cells end up in a single (stationary) cellular state. Several multistable regimes are observed along this route to oscillation death.