Phage-mediated intercellular CRISPRi for biocomputation in bacterial consortia - Archive ouverte HAL
Article Dans Une Revue Nucleic Acids Research Année : 2024

Phage-mediated intercellular CRISPRi for biocomputation in bacterial consortia

Résumé

Coordinated actions of cells in microbial communities and multicellular organisms enable them to perform complex tasks otherwise difficult for single cells. This has inspired biological engineers to build cellular consortia for larger circuits with improved functionalities while implementing communication sy stems f or coordination among cells. Here, w e in v estigate the signalling dynamics of a phage-mediated synthetic DNA messaging system and couple it with CRISPR interference to build distributed circuits that perform logic gate operations in multicellular bacterial consortia. We find that growth phases of both sender and receiver cells, as well as resource competition between them, shape communication outcomes. L e v eraging the easy programmability of DNA messages, we build eight orthogonal signals and demonstrate that intercellular CRISPRi (i-CRISPRi) regulates gene expression across cells. Finally, we multiplex the i-CRISPRi system to implement se v eral multicellular logic gates that in v olv e up to se v en cells and tak e up to three inputs simultaneously, with single-and dual-rail encoding: NOT, YES, AND and AND-AND-NOT. The communication system developed here lays the groundwork for implementing complex biological circuits in engineered bacterial communities, using phage signals for communication.
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Dates et versions

hal-04857279 , version 1 (28-12-2024)

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Abhinav Pujar, Amit Pathania, Corbin Hopper, Amir Pandi, Cristian Ruiz Calderón, et al.. Phage-mediated intercellular CRISPRi for biocomputation in bacterial consortia. Nucleic Acids Research, In press, pp.gkae1256. ⟨10.1093/nar/gkae1256⟩. ⟨hal-04857279⟩
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