McVeigh, Paul M. McCammick, Erin McCusker, Paul Morphew, Russell M. Mousley, Angela Abidi, Abbas M. Saifullah, Khalid Muthusamy, Raman Gopalakrishnan, Ravikumar W. Spithill, Terry Dalton, John P. Brophy, Peter M. J. Marks, Nikki Maule, Aaron G. Persistence of transcript knockdown during maintenance <i>in vitro</i>. <p>Following exposure to target long double-stranded (ds)RNA (A, cathepsin B; B, cathepsin L; C, sigma-class glutathione transferase), newly-excysted juveniles (NEJs) were maintained <i>in vitro</i> for up to 25 days. NEJs were collected and assayed by quantitative PCR at 3, 9 and 25 days post dsRNA exposure. Target ΔΔCt (Y-axes) represents ratio of abundance of target transcript to a reference gene (glyceraldehyde phosphate dehydrogenase, GAPDH), in treated samples, relative to the abundance of those transcripts in untreated samples. Statistical significances are indicated relative to effects of negative control dsRNA (dsCTRL, complementary to neomycin phosphotransferase). dsCTRL treatments were performed in parallel with all experimental treatments. Experiments were repeated ≥3 times, employing 20–30 flukes per replicate. *, P<0.05; **, P<0.01; ***, P<0.001. Symbols represent mean±SEM.</p> RNA interference;virulence genes;control target validation;27 nt;RNAi penetrance;dsRNA exposure;RNAi persistence;transcriptional suppression;Fhe σGST;anthelmintic resistance;Juvenile Fasciola spp;protein knockdown dynamics;Cysteine proteases;4 h exposure;probe RNAi dynamics;Gene function;Vitro BackgroundFasciola spp;cathepsin L;Fasciola gigantica juveniles;liver fluke NEJs;Fhe σGST FindingsRobust transcriptional;Vivo Studies;knockdown dynamics;siRNA triggers;protein suppression;liver fluke;Gene silencing;RNAi Dynamics;standardised RNAi protocols;Fasciola hepatica;FheCatL protein suppression;Liver Flukes;21 days;liver fluke cause 2014-09-25
    https://plos.figshare.com/articles/figure/_Persistence_of_transcript_knockdown_during_maintenance_in_vitro_/1182379
10.1371/journal.pntd.0003185.g005