Adverse effects induced by short hairpin RNA expression in porcine fetal fibroblasts.
RNA interference is a recent, gene silencing technique that could be extremely valuable in studying gene function, treating diseases, and developing novel animal models for human diseases. Here, we investigated the feasibility of applying shRNA-mediated RNA interference in fetal fibroblasts for silencing of the myostatin gene and investigate adverse effects of RNAi. We report that up to 97% silencing of myostatin mRNA was achieved using shRNA constructs in transiently and stably transfected fetal fibroblasts (p<0.05). At the same time we also demonstrate that high level of shRNA expression resulted in 10- to 1000-fold induction of interferon responsive genes (OAS1, IFN-beta) (p<0.05). In addition we also report novel adverse effect of shRNA expression in stably transfected cells-interference with microRNA processing/transport which led to 500-fold increase in the level of miR21 precursors (p<0.05). Reduction of these side effects will be essential to obtain long term stable RNAi silencing.
Stewart
Candace K
CK
Department of Animal and Poultry Science, University of Guelph, 50 Stone Road, Guelph, Ont., Canada N1G 2W1.
Li
Julang
J
Golovan
Serguei P
SP
D015088:2',5'-Oligoadenylate Synthetase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D005333:Fetus / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D007371:Interferon-gamma / Q000235:genetics / Q000378:metabolism; D007372:Interferons / Q000494:pharmacology; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D055435:Myostatin; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D034421:Sus scrofa; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors* / Q000235:genetics
C525911:MSTN protein, human; D035683:MicroRNAs; D055435:Myostatin; D034741:RNA, Small Interfering; D016212:Transforming Growth Factor beta; D007371:Interferon-gamma; D007372:Interferons; D015088:2',5'-Oligoadenylate Synthetase