Targeted delivery of diverse biomolecules with engineered bacterial nanosyringes

The Photorhabdus virulence cassette (PVC) is a microbial nanosyringe that can be engineered to deliver protein cargos into human cells. Here, we modify PVCs to incorporate exogenous cargos and targeting moieties in vitro. We show this method, termed SPEAR, enables loading of new types of cargo--including folded ribonucleoproteins (RNPs) and ssDNA--and targeting of defined cell types both in vitro and in vivo.