Multistable and dynamic CRISPRi-based synthetic circuits

Javier Santos-Moreno and Yolanda Schaerli

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Oct 15, 2019
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Received Date: 7th October 19

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, raising the question of whether CRISPRi is widely applicable for synthetic circuit design. Here we use CRISPRi to build prominent synthetic gene circuits that accurately govern temporal and spatial gene expression in Escherichia coli. We report the first-ever CRSPRi oscillator (“CRISPRlator”), bistable network and stripe pattern-forming incoherent feed-forward loop (IFFL). Our circuit designs, conceived to feature high predictability and orthogonality and low metabolic burden and context-dependency, allowed us to achieve robust circuit behaviors (e.g. synchronous oscillations) and to expand the IFFL into a twice as complex, two-stripe patterning system. 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.

Read in full at bioRxiv.

This is an abstract of a preprint hosted on an independent third party site. It has not been peer reviewed but is currently under consideration at Nature Communications.

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