Engineering Adhesion to Target Cells.

is a plant pathogen commonly repurposed for genetic modification of crops. Despite its versatility, it remains inefficient at transferring DNA to many hosts, including to animal cells. Like many pathogens, physical contact between and host cells promotes infection efficacy. Thus, improving the strength and specificity of to target cells has the potential for enhancing DNA transfer for biotechnological and therapeutic purposes. Here, we demonstrate a methodology for engineering genetically encoded exogeneous adhesins at the surface of . We identified an autotransporter gene we named Aat that is predicted to show canonical β-barrel and passenger domains. We engineered the β-barrel scaffold and linker (Aat) to display synthetic adhesins susceptible to rewire to alternative host targets. As a proof of concept, we leveraged the versatility of a VHH domain to rewire adhesion to yeast and mammalian hosts displaying a GFP target receptor. Finally, to demonstrate how synthetic adhesion can improve transfer to host cells, we showed improved protein translocation into HeLa cells using a sensitive split luciferase reporter system. Engineering adhesion has therefore a strong potential in generating complex heterogeneous cellular assemblies and in improving DNA transfer efficiency against non-natural hosts.