A novel eukaryote-like CRISPR activation tool in bacteria: features and capabilities

CRISPR activation (CRISPRa) in bacteria is an attractive method for programmable gene activation. Recently, we reported a eukaryote-like, σ54-dependent CRISPRa system. It exhibits high dynamic ranges and permits flexible target site selection. Here, we present an overview of the existing strategies of CRISPRa in bacteria, and introduce the characteristics and design principles of our CRISPRa system. We discuss possible scenarios for applying the eukaryote-like CRISPRa system with corresponding suggestions for performance optimization and future functional expansion. We envision the new eukaryote-like CRISPRa system enabling novel designs in multiplexed gene regulation and promote research in the σ54-dependent gene regulatory networks among a variety of biotechnology relevant or disease-associated bacterial species.