Adverse effects induced by short hairpin RNA expression in porcine fetal fibroblasts

• Published in: Biochemical and biophysical research communications 2008-05, Vol.370 (1), p.113-117
• Main Authors: Stewart, Candace K., Li, Julang, Golovan, Serguei P.
• Format: Article
• Language: English
• Subjects: 2',5'-Oligoadenylate Synthetase - genetics; 2',5'-Oligoadenylate Synthetase - metabolism; Animals; Cells, Cultured; Fetus - cytology; Fetus - metabolism; Fibroblasts - drug effects; Fibroblasts - metabolism; Interferon; Interferon-gamma - genetics; Interferon-gamma - metabolism; Interferons - pharmacology; MicroRNA; MicroRNAs - genetics; MicroRNAs - metabolism; Myostatin; RNA Interference; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; shRNA; Sus scrofa; Transfection; Transforming Growth Factor beta - antagonists & inhibitors; Transforming Growth Factor beta - genetics
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2008.03.041

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-β) ( 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.