Reho Guillaume, Menger Yannick, Lelièvre Vincent, Cadiou Hervé
UPR 3212, Institut des Neurosciences Cellulaires et Intégratives, Centre National de la Recherche Scientifique, Université de Strasbourg, Strasbourg, France.
UMR 7364, Laboratoire des Neurosciences Cognitives et Adaptatives, Centre National de la Recherche Scientifique, Université de Strasbourg, Strasbourg, France.
Front Neurosci. 2025 Apr 30;19:1546196. doi: 10.3389/fnins.2025.1546196. eCollection 2025.
RNA interference (RNAi) is a powerful tool to knock down the expression of genes of interests. In planarians, a popular animal model to study development and regeneration processes, RNAi is easily set up by feeding the animals double-stranded RNA (dsRNA). However, there is no consensus in the literature on the amount of dsRNA needed to efficiently knock down gene expression, nor on the lasting effect of this knockdown. Here, we exposed the worms to two RNAi protocols, either feeding them dsRNA only once or three times in the span of a week. To observe the gradual loss and retrieval of nociceptive phenotypes, we exposed the worms to Allyl Isothiocyanate (AITC), an irritant and TRPA1 receptor agonist, while we knocked down the expression of the TRPA1 receptor and performed behavioral assessments over 11 weeks. We showed that feeding planarians once was sufficient to induce similar phenotypes as feeding them three times, that also lasted as long. These insights are useful to refine RNAi protocol timelines and may save some valuable resources.
RNA干扰（RNAi）是一种强大的工具，可用于降低目标基因的表达。在涡虫（一种用于研究发育和再生过程的常用动物模型）中，通过给动物喂食双链RNA（dsRNA）可轻松建立RNAi。然而，文献中对于有效降低基因表达所需的dsRNA量以及这种敲低的持久效果尚无共识。在此，我们让涡虫接受两种RNAi方案，即在一周内只给它们喂食一次dsRNA或三次dsRNA。为了观察伤害感受性表型的逐渐丧失和恢复，我们让涡虫接触异硫氰酸烯丙酯（AITC，一种刺激物和TRPA1受体激动剂），同时敲低TRPA1受体的表达，并在11周内进行行为评估。我们发现，给涡虫喂食一次足以诱导出与喂食三次相似的表型，且持续时间相同。这些见解有助于优化RNAi方案的时间安排，并可能节省一些宝贵的资源。
