Transfection of siRNA into Brachionus plicatilis (Rotifera)

摘要: Transfection experiments have played a prominent role in functional genomics, being used to probe phenotypes with reporters and knockouts techniques like RNAi, gene over-expression studies, the creation of transgenic animals. Rotifers are not amenable many classic genetic manipulations, hence been model system genomics revolution. However, no one has explored whether rotifers susceptible expression knock-out via transfection, establishment rotifer transcriptome sequencing project, an increasing amount sequence data is available define target genes for manipulation. As first step towards transfection rotifers, we describe our attempt transfect double-stranded, fluorescently-labeled siRNA into resting eggs, hatched amictic eggs varying levels success. was successful when (rather than or eggs) were directly treated, possibly treated as eggs. Highest (as determined by significant fluorescence pseudocoelom) observed fed that 2 h old at time treatment, which involved combining lipofection electroporation methods transfection. Successful these animals now provides possible mechanism exploration function.