A Dueling-Competent Signal-Sensing Module Guides Precise Delivery of Cargo Proteins into Target Cells by Engineered Pseudomonas aeruginosa

To recognize and manipulate a specific microbe of a crowded community is a highly challenging task in synthetic biology. Here we introduce a highly selective protein delivery platform, termed DUEC, which responds to direct contact of attacking cells by engineering the tit-for-tat/dueling response of H1-T6SS (type VI secretion system) in Pseudomonas aeruginosa. Using a Cre-recombinase-dependent reporter, we screened H1-T6SS-secreted substrates and developed Tse6N as the most effective secretion tag for Cre delivery. DUEC cells can discriminately deliver the Tse6N-Cre cargo into the cytosol of T6SS+ but not T6SS- Vibrio cholerae cells. DUEC could also deliver a nuclease cargo, Tse6N-NucSe1, to selectively kill provoking cells in a mixed community. These data demonstrate that the DUEC cell not only is a prototypical physical-contact sensor and delivery platform but also may be coupled with recombination-based circuits with the potential for complex tasks in mixed microbial communities.

Automated summary

We have developed a highly selective protein delivery platform, DUEC, which responds to direct contact of attacking cells by engineering the tit-for-tat/dueling response of H1-T6SS in Pseudomonas aeruginosa. By screening H1-T6SS-secreted substrates, we have created Tse6N as an effective secretion tag for delivering Cre. DUEC cells can selectively deliver the Tse6N-Cre cargo into T6SS+ but not T6SS- Vibrio cholerae cells, and can also deliver a nuclease cargo to selectively kill provoking cells in a mixed community.