title: MicroRNA-mediated myostatin silencing in caprine fetal fibroblasts.
abstract: Myostatin functions as a negative regulator of skeletal muscle growth by suppressing proliferation and differentiation of myoblasts. Dysfunction of the myostatin gene, either due to natural mutation or genetic manipulations such as knockout or knockdown, has been reported to increase muscle mass in mammalian species. RNA interference (RNAi) mediated by microRNAs (miRNAs) is a promising method for gene knockdown studies. In the present study, transient and stable silencing of the myostatin gene in caprine fetal fibroblasts (CFF) was evaluated using the two most effective constructs selected from four different miRNA expression constructs screened in 293FT cells. Using these two miRNA constructs, we achieved up to 84% silencing of myostatin mRNA in transiently transfected CFF cells and up to 31% silencing in stably transfected CFF cells. Moreover, off-target effects due to induction of interferon (IFN) response genes, such as interferon beta (IFN-β) and 2'-5'-oligoadenylate synthetase 2 (OAS2), were markedly fewer in stably transfected CFF cells than in transiently transfected cells. Stable expression of anti-myostatin miRNA with minimal induction of interferon shows great promise for increasing muscle mass in transgenic goats.
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, PR China; Jiangsu Engineering Technology Research Center of Meat Sheep & Goat Industry, Nanjing Agricultural University, Nanjing, PR China.
authors.forename: Bushuai
authors.initials: B
authors.lastname: Zhong
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, PR China; Jiangsu Engineering Technology Research Center of Meat Sheep & Goat Industry, Nanjing Agricultural University, Nanjing, PR China.
authors.forename: Yanli
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, PR China.
authors.forename: Yibo
authors.initials: Y
authors.lastname: Yan
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, PR China.
authors.forename: Ziyu
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Jiangsu Engineering Technology Research Center of Meat Sheep & Goat Industry, Nanjing Agricultural University, Nanjing, PR China.
authors.forename: Shijia
authors.initials: S
authors.lastname: Ying
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, PR China.
authors.forename: Mingrui
authors.initials: M
authors.lastname: Huang
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, PR China; Jiangsu Engineering Technology Research Center of Meat Sheep & Goat Industry, Nanjing Agricultural University, Nanjing, PR China.
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
mesh_terms: D000818:Animals; D005347:Fibroblasts / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D006041:Goats; D057809:HEK293 Cells; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D055435:Myostatin / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference
chemical_list: D035683:MicroRNAs; D055435:Myostatin