Layering genetic circuits to build a single cell, bacterial half adder.

BACKGROUND: Gene regulation in biological systems is impacted by the cellular and genetic context-dependent effects of the biological parts which comprise the circuit. Here, we have sought to elucidate the limitations of engineering biology from an architectural point of view, with the aim of compiling a set of engineering solutions for overcoming failure modes during the development of complex, synthetic genetic circuits. RESULTS: Using a synthetic biology approach that is supported by computational modelling and rigorous characterisation, AND, OR and NOT biological logic gates were layered in both parallel and serial arrangements to generate a repertoire of Boolean operations that include NIMPLY, XOR, half adder and half subtractor logics in a single cell. Subsequent evaluation of these near-digital biological systems revealed critical design pitfalls that triggered genetic context-dependent effects, including 5' UTR interferences and uncontrolled switch-on behaviour of the supercoiled σ54 promoter. In particular, the presence of seven consecutive hairpins immediately downstream of the promoter transcription start site severely impeded gene expression. CONCLUSIONS: As synthetic biology moves forward with greater focus on scaling the complexity of engineered genetic circuits, studies which thoroughly evaluate failure modes and engineering solutions will serve as important references for future design and development of synthetic biological systems. This work describes a representative case study for the debugging of genetic context-dependent effects through principles elucidated herein, thereby providing a rational design framework to integrate multiple genetic circuits in a single prokaryotic cell.

A novel neural-inspired learning algorithm with application to clinical risk prediction.

Clinical risk prediction - the estimation of the likelihood an individual is at risk of a disease - is a coveted and exigent clinical task, and a cornerstone to the recommendation of life saving management strategies. This is especially important for individuals at risk of cardiovascular disease (CVD) given the fact that it is the leading causes of death in many developed counties. To this end, we introduce a novel learning algorithm - a key factor that influences the performance of machine learning-based prediction models - and utilities it to develop CVD risk prediction tool. This novel neural-inspired algorithm, called the Artificial Neural Cell System for classification (ANCSc), is inspired by mechanisms that develop the brain and empowering it with capabilities such as information processing/storage and recall, decision making and initiating actions on external environment. Specifically, we exploit on 3 natural neural mechanisms responsible for developing and enriching the brain - namely neurogenesis, neuroplasticity via nurturing and apoptosis - when implementing ANCSc algorithm. Benchmark testing was conducted using the Honolulu Heart Program (HHP) dataset and results are juxtaposed with 2 other algorithms - i.e. Support Vector Machine (SVM) and Evolutionary Data-Conscious Artificial Immune Recognition System (EDC-AIRS). Empirical experiments indicate that ANCSc algorithm (statistically) outperforms both SVM and EDC-AIRS algorithms. Key clinical markers identified by ANCSc algorithm include risk factors related to diet/lifestyle, pulmonary function, personal/family/medical history, blood data, blood pressure, and electrocardiography. These clinical markers, in general, are also found to be clinically significant - providing a promising avenue for identifying potential cardiovascular risk factors to be evaluated in clinical trials.

A biological continuum based approach for efficient clinical classification.

Clinical feature selection problem is the task of selecting and identifying a subset of informative clinical features that are useful for promoting accurate clinical diagnosis. This is a significant task of pragmatic value in the clinical settings as each clinical test is associated with a different financial cost, diagnostic value, and risk for obtaining the measurement. Moreover, with continual introduction of new clinical features, the need to repeat the feature selection task can be very time consuming. Therefore to address this issue, we propose a novel feature selection technique for diagnosis of myocardial infarction - one of the leading causes of morbidity and mortality in many high-income countries. This method adopts the conceptual framework of biological continuum, the optimization capability of genetic algorithm for performing feature selection and the classification ability of support vector machine. Together, a network of clinical risk factors, called the biological continuum based etiological network (BCEN), was constructed. Evaluation of the proposed methods was carried out using the cardiovascular heart study (CHS) dataset. Results demonstrate a significant speedup of 4.73-fold can be achieved for the development of MI classification model. The key advantage of this methodology is the provision of a reusable (feature subset) paradigm for efficient development of up-to-date and efficacious clinical classification models.

Building the UK's industrial base in engineering biology.

The paper describes the strategy and components that have been put in place to build the UK's research and industrial base in Engineering Biology. The initial section of the paper provides a brief historical overview of the development of the field in the United Kingdom. This comprised, principally, a major report by the Royal Academy of Engineering and a strategic roadmap for synthetic biology, together with the establishment of six new synthetic biology research centres, a national centre for the industrial translation of synthetic biology and five biofoundries. The next section of the paper describes the UK government's policy for the field. Important elements of the implementation of the policy comprises people, Infrastructure, Business Environment and place. In this context, a number of important areas are addressed-including industrial translation; building an expert workforce and nucleating, incubating and accelerating a new engineering biology industry in the United Kingdom. The final portion of the paper addresses the author's view of the way forward. This comprises placing the development of the field, both nationally and internationally, in the context of the development of the Bioeconomy and Climate Change. The final section of the text addresses a specific strategic approach and the implications for the United Kingdom in relation to the development of its industrial base in Engineering Biology.

Build a Sustainable Vaccines Industry with Synthetic Biology.

The vaccines industry has not changed appreciably in decades regarding technology, and has struggled to remain viable, with large companies withdrawing from production. Meanwhile, there has been no let-up in outbreaks of viral disease, at a time when the biopharmaceuticals industry is discussing downsizing. The distributed manufacturing model aligns well with this, and the advent of synthetic biology promises much in terms of vaccine design. Biofoundries separate design from manufacturing, a hallmark of modern engineering. Once designed in a biofoundry, digital code can be transferred to a small-scale manufacturing facility close to the point of care, rather than physically transferring cold-chain-dependent vaccine. Thus, biofoundries and distributed manufacturing have the potential to open up a new era of biomanufacturing, one based on digital biology and information systems. This seems a better model for tackling future outbreaks and pandemics.

Miniaturisation of high-throughput plasmid DNA library preparation for next-generation sequencing using multifactorial optimisation.

High-throughput preparation of plasmid DNA libraries for next-generation sequencing (NGS) is an important capability for molecular biology laboratories. In particular, it is an essential quality control (QC) check when large numbers of plasmid variants are being generated. Here, we describe the use of the Design of Experiments (DOE) methodology to optimise the miniaturised preparation of plasmid DNA libraries for NGS, using the Illumina® Nextera XT technology and the Labcyte Echo® acoustic liquid dispensing system. Furthermore, we describe methods which can be implemented as a QC check for identifying the presence of genomic DNA (gDNA) in plasmid DNA samples and the subsequent shearing of the gDNA, which otherwise prevents the acoustic transfer of plasmid DNA. This workflow enables the preparation of plasmid DNA libraries which yield high-quality sequencing data.

Author Correction: Building a global alliance of biofoundries.

The original version of this Comment contained errors in the legend of Figure 2, in which the locations of the fifteenth and sixteenth GBA members were incorrectly given as '(15) Australian Genome Foundry, Macquarie University; (16) Australian Foundry for Advanced Biomanufacturing, University of Queensland.'. The correct version replaces this with '(15) Australian Foundry for Advanced Biomanufacturing (AusFAB), University of Queensland and (16) Australian Genome Foundry, Macquarie University'. This has been corrected in both the PDF and HTML versions of the Comment.

Single-trial EEG source reconstruction for brain-computer interface.

A new way to improve the classification rate of an EEG-based brain-computer interface (BCI) could be to reconstruct the brain sources of EEG and to apply BCI methods to these derived sources instead of raw measured electrode potentials. EEG source reconstruction methods are based on electrophysiological information that could improve the discrimination between BCI tasks. In this paper, we present an EEG source reconstruction method for BCI. The results are compared with results from raw electrode potentials to enable direct evaluation of the method. Features are based on frequency power change and Bereitschaft potential. The features are ranked with mutual information before being fed to a proximal support vector machine. The dataset IV of the BCI competition II and data from four subjects serve as test data. Results show that the EEG inverse solution improves the classification rate and can lead to results comparable to the best currently known methods.

Wavelet-based de-noising algorithm for images acquired with parallel magnetic resonance imaging (MRI).

Wavelet-based de-noising has been shown to improve image signal-to-noise ratio in magnetic resonance imaging (MRI) while maintaining spatial resolution. Wavelet-based de-noising techniques typically implemented in MRI require that noise displays uniform spatial distribution. However, images acquired with parallel MRI have spatially varying noise levels. In this work, a new algorithm for filtering images with parallel MRI is presented. The proposed algorithm extracts the edges from the original image and then generates a noise map from the wavelet coefficients at finer scales. The noise map is zeroed at locations where edges have been detected and directional analysis is also used to calculate noise in regions of low-contrast edges that may not have been detected. The new methodology was applied on phantom and brain images and compared with other applicable de-noising techniques. The performance of the proposed algorithm was shown to be comparable with other techniques in central areas of the images, where noise levels are high. In addition, finer details and edges were maintained in peripheral areas, where noise levels are low. The proposed methodology is fully automated and can be applied on final reconstructed images without requiring sensitivity profiles or noise matrices of the receiver coils, therefore making it suitable for implementation in a clinical MRI setting.

Addressing the future of clinical information systems--Web-based multilayer visualization.

This paper addresses some key issues relating to the development of new technology for clinical information systems (CIS) in relation to imaging and visualizing data. With the increasing importance of molecular and cellular biology, a new type of medicine, molecular based medicine, is now developing. This will significantly alter the way in which medicine is practiced. The view is presented that CIS will need to operate seamlessly across the Biological Continuum, i.e., the hierarchy of the human organism comprising systems, viscera, tissue, cells, proteins, and genes. We propose a multilayered visualization interface, which operates across the Biological Continuum, based on Web-based technology. A visualization interface package for two-dimensional and three-dimensional image data at the visceral and cellular levels is described. Two application examples are presented: 1) MR knee images, at the visceral level and 2) endothelial nuclei images, acquired from confocal laser microscopy, at the cellular level.