Engineering principles of synthetic biochemical oscillators with negative cyclic feedback

This paper analyzes oscillatory dynamics of a class of cyclic gene regulatory networks and provides engineering principles for the synthesis of biochemical oscillators. We first review previous results that the oscillatory parameter regime of the gene regulatory circuits can be rigorously explored by the local stability analysis of a unique equilibrium. The local stability analysis then leads to the first engineering principle that the circuit components, or genes, should be chosen so that the kinetic profiles of the circuit components are similar to each other. Using a homogeneous oscillator model, we further discuss design strategies to reduce the cell-to-cell variability of the oscillators that is caused by intrinsic noise

Collective oscillation period of inter-coupled biological negative cyclic feedback oscillators

A number of biological rhythms originate from networks comprised of multiple cellular oscillators. But analytical results are still lacking on the collective oscillation period of inter-coupled gene regulatory oscillators, which, as has been reported, may be different from that of an autonomous oscillator. Based on cyclic feedback oscillators, we analyze the collective oscillation pattern of coupled cellular oscillators. First we give a condition under which the oscillator network exhibits oscillatory and synchronized behavior. Then we estimate the collective oscillation period based on a novel multivariable harmonic balance technique. Analytical results are derived in terms of biochemical parameters, thus giving insight into the basic mechanism of biological oscillation and providing guidance in synthetic biology design.Comment: arXiv admin note: substantial text overlap with arXiv:1203.125

Biological Oscillators in Nanonetworks-Opportunities and Challenges.

One of the major issues in molecular communication-based nanonetworks is the provision and maintenance of a common time knowledge. To stay true to the definition of molecular communication, biological oscillators are the potential solutions to achieve that goal as they generate oscillations through periodic fluctuations in the concentrations of molecules. Through the lens of a communication systems engineer, the scope of this survey is to explicitly classify, for the first time, existing biological oscillators based on whether they are found in nature or not, to discuss, in a tutorial fashion, the main principles that govern the oscillations in each oscillator, and to analyze oscillator parameters that are most relevant to communication engineer researchers. In addition, the survey highlights and addresses the key open research issues pertaining to several physical aspects of the oscillators and the adoption and implementation of the oscillators to nanonetworks. Moreover, key research directions are discussed

Design and implementation of a mammalian synthetic gene oscillator

The core goal of synthetic biology as a discipline is to design, develop and characterize biological parts in order to precisely control cellular behaviour. Much of the research in this field has been focused on the development of gene regulatory networks, namely switches and oscillators. The study of synthetic gene oscillators has attracted significant attention in the past decade due to their intriguing dynamics and relevance in controlling inflammatory, metabolic and circadian signalling pathways. Additionally, the precise expression dynamics and molecular mechanisms that underlie the mammalian circadian clock structure are not fully understood. The work presented herein regards the design and implementation of a tuneable mammalian synthetic gene oscillator with a novel biological structure. To this end, an approach based on a combination of in silico design and in vivo part validation, in conjunction with a comparative analysis of previously implemented synthetic gene oscillators, was taken when assembling the proposed system. The topology of the system relies on a delayed negative feedback loop, consisting of the coupled regulatory activities of the transcription regulators LacI, tTA, and Gal4. The numerical solution and stability analysis of an ODE-based model describing the dynamics of the system are indicative that the proposed system is capable of generating sustained oscillations across a wide range of parameter values. The biological parts that comprise the system have been monitored and validated in HEK293T cells through time-lapse fluorescence microscopy and image analysis. The in vivo performance of the proposed mammalian synthetic gene oscillator was also assessed in the HEK293T cell line, and monitored using time-lapse fluorescence microscopy. Damped fluorescence oscillations were observed: these could be tuned by a differential IPTG concentration gradient and abolished by doxycycline. The proposed mammalian synthetic gene oscillator provides valuable insight into the gene expression regulatory processes leading to oscillatory behaviour, and has the potential to foster progress in future synthetic biology-based therapies.Open Acces

Reliability of Transcriptional Cycles and the Yeast Cell-Cycle Oscillator

A recently published transcriptional oscillator associated with the yeast cell cycle provides clues and raises questions about the mechanisms underlying autonomous cyclic processes in cells. Unlike other biological and synthetic oscillatory networks in the literature, this one does not seem to rely on a constitutive signal or positive auto-regulation, but rather to operate through stable transmission of a pulse on a slow positive feedback loop that determines its period. We construct a continuous-time Boolean model of this network, which permits the modeling of noise through small fluctuations in the timing of events, and show that it can sustain stable oscillations. Analysis of simpler network models shows how a few building blocks can be arranged to provide stability against fluctuations. Our findings suggest that the transcriptional oscillator in yeast belongs to a new class of biological oscillators

Rapid cell-free forward engineering of novel genetic ring oscillators

While complex dynamic biological networks control gene expression in all living organisms, the forward engineering of comparable synthetic networks remains challenging. The current paradigm of characterizing synthetic networks in cells results in lengthy design-build-test cycles, minimal data collection, and poor quantitative characterization. Cell-free systems are appealing alternative environments, but it remains questionable whether biological networks behave similarly in cell-free systems and in cells. We characterized in a cell-free system the 'repressilator,' a three-node synthetic oscillator. We then engineered novel three, four, and five-gene ring architectures, from characterization of circuit components to rapid analysis of complete networks. When implemented in cells, our novel 3-node networks produced population-wide oscillations and 95% of 5-node oscillator cells oscillated for up to 72 hours. Oscillation periods in cells matched the cell-free system results for all networks tested. An alternate forward engineering paradigm using cell-free systems can thus accurately capture cellular behavior

Designer Gene Networks: Towards Fundamental Cellular Control

The engineered control of cellular function through the design of synthetic genetic networks is becoming plausible. Here we show how a naturally occurring network can be used as a parts list for artificial network design, and how model formulation leads to computational and analytical approaches relevant to nonlinear dynamics and statistical physics.Comment: 35 pages, 8 figure

Synthetic Modeling and Mechanistic Account: Material Recombination and Beyond

Recently, Bechtel and Abrahamsen have argued that mathematical models study the dynamics of mechanisms by recomposing the components and their operations into an appropriately organized system. We will study this claim through the practice of combinational modeling in circadian clock research. In combinational modeling experiments on model organisms and mathematical/computational models are combined with a new type of model—a synthetic model. While we appreciate Bechtel and Abrahamsen’s point that mathematical/computational models are used to provide dynamic mechanistic explanations, we think that the strategy of recomposition is more complicated than what Bechtel and Abrahamsen indicate. Moreover, as we will show, synthetic modeling as a kind of material recomposition strategy also points beyond the mechanistic paradigm