Gene expression is unpredictable, noisy and prone to perturbations, thus posing a challenge towards designing reliable and functional systems. One of the most decisive factors affecting the functionality of synthetic biological systems is gene copy number variation, that gives rise to variability in gene expression.
We provide Galene, a genetic toolbox that enhances system stability by disassociating gene expression from copy number and provides translational control of protein production.

The former is achieved by implementing a type I incoherent feed-forward loop network motif that consists of a regulatory element which compensates for changes in copy number, thus stabilizing the promoter. Through model-driven feedback, stabilization by utilizing 2 distinct systems, each with its own advantages and appropriate uses. CRISPR interference, TAL Effectors.

In addition, we designed and simulated a 3rd stabilization system, based on transcriptional attenuator-interacting sRNAs

Going one step further, we implemented a theophylline aptamer that regulates translation and allows for the induction at the desired expression level.
This year, our Human Practice efforts were divided into 4 different categories, each of them affecting our project and ourselves in a different way. We focused on: taking into serious consideration ethical issues regarding our project, reaching out to field experts, interacting with people from the industry in order to investigate various possibilities and options for our project, and communicating science, synthetic biology and our project across people of different ages, scientific and cultural backgrounds. As a highlight of our engagement activities, we organized a series of synthetic biology events in the Balkan Peninsula.