title: RNA interference by feeding in vitro-synthesized double-stranded RNA to planarians: methodology and dynamics.
abstract: BACKGROUND
  The ability to assess gene function is essential for understanding biological processes. Currently, RNA interference (RNAi) is the only technique available to assess gene function in planarians, in which it has been induced by means of injection of double-stranded RNA (dsRNA), soaking, or ingestion of bacteria expressing dsRNA.
  
  
  RESULTS
  We describe a simple and robust RNAi protocol, involving in vitro synthesis of dsRNA that is fed to the planarians. Advantages of this protocol include the ability to produce dsRNA from any vector without subcloning, resolution of ambiguities in quantity and quality of input dsRNA, as well as time and ease of application. We have evaluated the logistics of inducing RNAi in planarians using this methodology in careful detail, from the ingestion and processing of dsRNA in the intestine, to timing and efficacy of knockdown in neoblasts, germline, and soma. We also present systematic comparisons of effects of amount, frequency, and mode of dsRNA delivery.
  
  
  CONCLUSIONS
  This method gives robust and reproducible results and is amenable to high-throughput studies. Overall, this RNAi methodology provides a significant advance by combining the strengths of current protocols available for dsRNA delivery in planarians and has the potential to benefit RNAi methods in other systems.
authors: Howard Hughes Medical Institute and Department of Cell and Developmental Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA. rouhana@life.illinois.edu
authors: Labib
authors: L
authors: Rouhana
authors: Jennifer A
authors: JA
authors: Weiss
authors: David J
authors: DJ
authors: Forsthoefel
authors: Hayoung
authors: H
authors: Lee
authors: Ryan S
authors: RS
authors: King
authors: Takeshi
authors: T
authors: Inoue
authors: Norito
authors: N
authors: Shibata
authors: Kiyokazu
authors: K
authors: Agata
authors: Phillip A
authors: PA
authors: Newmark
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics; D015509:Developmental Biology / Q000379:methods; D018507:Gene Expression Regulation, Developmental; D005821:Genetic Techniques; D005822:Genetic Vectors; D010641:Phenotype; D010932:Planarians / Q000235:genetics*; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics*; D015203:Reproducibility of Results
chemical_list: D012330:RNA, Double-Stranded