Engineering CRISPR-based Synthetic Circuits in Bacteria
Synthetic gene circuits allow us to govern cell behaviour in a programmable manner for user-defined tasks. Transcription factors constitute the ‘classic’ tool for synthetic circuit construction but some of their inherent constraints, such as insufficient modularity, orthogonality and programmability, limit progress in realizing more ambitious designs. CRISPR (clustered regularly interspaced short palindromic repeats) technology offers new and powerful possibilities for synthetic circuit design.
I will present our efforts in engineering CRISPR-based synthetic circuits in bacteria. First, I will show how we used CRISPR interference to build synthetic circuits such as an oscillator (“CRISPRlator”) and incoherent feed-forward loops in Escherichia coli. The ease of building such circuits let us to explore the genotype-phenotype map of small network motifs and gain insights into their evolution and function. I then will present that the CRISPRlator also functions in Streptococcus pneumoniae and how we used it to demonstrate that variation in capsule production is beneficial for pneumococcal fitness in traits associated with pathogenesis. Finally, I will present how we can combine CRISPRi with CRISPR activation (CRISPRa) in the same circuits.