Multistable and dynamic CRISPRi-based synthetic circuits

Javier Santos-Moreno, Eve Tasiudi, Joerg Stelling and Yolanda Schaerli ()
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Javier Santos-Moreno: University of Lausanne
Eve Tasiudi: ETH Zurich and SIB Swiss Institute of Bioinformatics
Joerg Stelling: ETH Zurich and SIB Swiss Institute of Bioinformatics
Yolanda Schaerli: University of Lausanne

Nature Communications, 2020, vol. 11, issue 1, 1-8

Abstract: Abstract Gene expression control based on CRISPRi (clustered regularly interspaced short palindromic repeats interference) has emerged as a powerful tool for creating synthetic gene circuits, both in prokaryotes and in eukaryotes; yet, its lack of cooperativity has been pointed out as a potential obstacle for dynamic or multistable synthetic circuit construction. Here we use CRISPRi to build a synthetic oscillator (“CRISPRlator”), bistable network (toggle switch) and stripe pattern-forming incoherent feed-forward loop (IFFL). Our circuit designs, conceived to feature high predictability and orthogonality, as well as low metabolic burden and context-dependency, allow us to achieve robust circuit behaviors in Escherichia coli populations. Mathematical modeling suggests that unspecific binding in CRISPRi is essential to establish multistability. Our work demonstrates the wide applicability of CRISPRi in synthetic circuits and paves the way for future efforts towards engineering more complex synthetic networks, boosted by the advantages of CRISPR technology.

Date: 2020
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DOI: 10.1038/s41467-020-16574-1

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