Protocol combining RNA interference and regeneration assays in planarian embryos

Planarian flatworms gradually acquire whole-body regeneration abilities during late embryonic and juvenile development. Here we show how to perturb gene function with RNA interference (RNAi) in S. polychroa embryos at developmental stages capable of wound healing following amputation. We describe embryo staging, embryo amputation and double-stranded RNA soaking, phenotype analysis, and techniques for qualitative and quantitative assessment of RNAi knock-down efficacy. 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 ¹.