PT  - JOURNAL ARTICLE
AU  - Javier Santos-Moreno
AU  - Eve Tasiudi
AU  - Joerg Stelling
AU  - Yolanda Schaerli
TI  - Multistable and dynamic CRISPRi-based synthetic circuits
AID  - 10.1101/756338
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 756338
4099  - http://biorxiv.org/content/early/2020/03/07/756338.short
4100  - http://biorxiv.org/content/early/2020/03/07/756338.full
AB  - 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 circuit construction. Here we use CRISPRi to build prominent synthetic gene circuits in Escherichia coli. We report the first-ever CRISPRi 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, allowed us to achieve robust circuit behaviors. 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.