A unified platform for experimental and computational biology

Abstract

In natural sciences, the correct engineering of a system’s chemical, biological and physical properties may allow it to sustain life. Bioengineering cells is probably one of the most complex challenges of biological research; yet, the little we do know about the nature of life is sufficient to guide scientific research, and to explore the elements beyond the apparent simple proliferation of living cells. Although Mendel first characterised the concept of genetic heredity over 150 years ago, we only recently became able to perform tailored genetic modification of living organisms. The development of digital technologies, in particular, has positively influenced the quality and reproducibility of experimental results emerging from biological assays. However, the use of any equipment may require the need for a specific expertise in order to perform a given experimental procedure. Therefore, multidisciplinary research can bring benefits to all fields of science by helping the development of analytical methods that cross the boundaries of individual disciplines. This emerges as a systematic view of scientific problems, and relies on the adequation and integration of results from different research areas. Nevertheless, there is a complex interface between hard sciences that often creates a gap between experimental and theoretical models. In this thesis, we explored synthetic biology approaches and created a unified platform to fill this gap. We propose the first barcoding platform (Bac2code) that allows the identification and the tracking of bacterial strains. In order to facilitate communication between researchers, we developed a barcode system in DNA that physically links bacteria to their genetic description. We designed DNA barcodes as bioorthogonal elements, elaborated a universal cloning strategy to integrate these sequences in Gram-negative and Gram-positive bacteria, and demonstrated their stability over time. Through a generic protocol, any barcoded strain can later be identified via a single sequencing read. With the engineering of a synthetic circuit library, we built a biorepository of genetic constructs for our barcoding platform. These biological devices were optimised based on the closest achievable interface between experimental biology and viii computational results. Following their characterisation, and in the context of intercellular communication, we studied the behaviour of small cohorts of bioengineered cells at the microscale in microfluidics. We pushed the biological and physical boundaries of engineering techniques to the maximum, in order to observe physiological changes between bacteria separated by distances down to 20µm. However, we also showed that we reached a technological barrier, where even the use of nanoscale features was found insufficient to maintain cells isolated under high cellular density. Yet, microfluidics remains a remarkable technology, and we propose the expansion of barcoding methods to automated systems, which would allow serial barcode integration and documentation retrieval at any one time. Here, we developed and tested a barcoding method to ensure the cohesion of experimental and computational biology resources. We demonstrated its use by the in vitro assembly and the in vivo or in silico characterisation of a series of genetic circuits via different techniques. The research output of this thesis is realised as a step forward in interdisciplinary studies, and is now being adapted to reach a larger community of users as a startup company