Cellulose, a ubiquitous polysaccharide with critical roles in life, provides structural integrity to cells in plants and aids biofilm formation in many bacteria. Although bacterial cellulose biosynthe...

Streptomycetes hold immense potential for discovering novel bioactive molecules for applications in medicine or sustainable agriculture. However, high-throughput exploration is hampered by the current Streptomyces genetic engineering methods that involve the manual design of complex experimental molecular biological engineering strategies for each targeted gene. Here, we introduce StreptoCAD, an open-source software toolbox that automates and streamlines the design of genome engineering strategies in Streptomyces, supporting various CRISPR-based and gene overexpression methods. Once initiated, StreptoCAD designs all necessary DNA primers and CRISPR guide sequences, simulates plasmid assemblies (cloning) and the resulting modification of the genomic target(s), and further summarizes the information needed for laboratory implementation and documentation. StreptoCAD currently offers six design workflows, including the construction of overexpression libraries, base-editing, including multiplexed CRISPR-BEST plasmid generation, and genome engineering using CRISPR-Cas9, CRISPR-Cas3, and CRISPRi systems. In addition to automating the design process, StreptoCAD further secures compliance with the FAIR principles, ensuring reproducibility and ease of data management via standardized output files. To experimentally demonstrate the design process and output of StreptoCAD, we designed and constructed a series of gene overexpression strains, and performed CRISPRi knockdowns in Streptomyces Gö40/10, underscoring the tool’s efficiency and user-friendliness.. This tool simplifies complex genetic engineering tasks and promotes collaboration through standardized workflows and design parameters. StreptoCAD is set to transform genome engineering in Streptomyces, making sophisticated genetic manipulations accessible for all and accelerating natural product discovery.

Lipoproteins are major cell-surface components in archaea, but their functions and the lipidation mechanisms are unclear. Here, Hong et al. identify two proteins required for attachment of proteins to unique archaeal membrane lipids via thioether bonds, and demonstrate their importance in archaeal physiology.

Common mycorrhizal networks (CMNs) facilitate chemical communication between plants. Zhang et al. demonstrate the transfer of jasmonic acid via CMN from Botrytis cinerea-infected donor plants to healt...

Granaticins, a class of bioactive benzoisochromanequinones (BIQs), are natural blue pigments with biological activities, including antibacterial prope…