Double‐stranded RNA (dsRNA) was synthesized as previously described (Rouhana et al, 2013 (link)). Briefly, in vitro transcription (IVT) templates were prepared by PCR amplification of cloned target sequences using primers with 5′ flanking T7 promoter sequences. dsRNA was synthesized using the TranscriptAid T7 High Yield Transcription Kit (CAT K0441; Thermo Scientific). Reactions were incubated overnight at 37°C and then supplemented with RNase‐free DNase for 30 min. RNA was purified by ethanol precipitation and finally resuspended in 25 μl of ddH2O. RNA was analyzed on 1% agarose gel and quantified by Qubit 4 (Thermo scientific) for validating a concentration higher than 5 μg/μl. Animals were starved for at least 7 days prior to RNAi experiments. Schmidtea mediterranea, asexual, were used for all experiments. Animals were fed with dsRNA mixed with beef liver twice a week as previously described (Wurtzel et al, 2017 (link)).