Protocol combining RNA interference and regeneration assays in planarian embryos

Planarians acquire regenerative abilities during late embryonic and juvenile development. Here, we present a protocol combining gene perturbation by RNA interference (RNAi) and regeneration assays in S. polychroa embryos. We describe steps for embryo staging, amputation, double-stranded RNA soaking, and phenotype analysis. This protocol is adaptable for use with embryonic stages amenable to ex vivo culturing and with other embryo-producing flatworm species. For complete details on the use and execution of this protocol, please refer to Booth et al.¹.