title: Effect of Co-transfection of Anti-myostatin shRNA Constructs in Caprine Fetal Fibroblast Cells.
abstract: Knockdown of myostatin gene (MSTN), transforming growth factor-β superfamily, and a negative regulator of the skeletal muscle growth, by RNA interference (RNAi), has been reported to increase muscle mass in mammals. The current study was aimed to cotransfect two anti-MSTN short hairpin RNA (shRNA) constructs in caprine fetal fibroblast cells for transient silencing of MSTN gene. In the present investigation, approximately 89% MSTN silencing was achieved in transiently transfected caprine fetal fibroblast cells by cotransfection of two best out of four anti-MSTN shRNA constructs. Simultaneously, we also monitored the induction of IFN responsive genes (IFN), pro-apoptotic gene (caspase3) and anti-apoptotic gene (MCL-1) due to cotransfection of different anti-MSTN shRNA constructs. We observed induction of 0.66-19.12, 1.04-4.14, 0.50-3.43, and 0.42-1.98 for folds IFN-β, OAS1, caspase3, and MCL-1 genes, respectively (p < 0.05). This RNAi based cotransfection method could provide an alternative strategy of gene knockout and develop stable caprine fetal fibroblast cells. Furthermore, these stable cells can be used as a cell donor for the development of transgenic cloned embryos by somatic cell nuclear transfer (SCNT) technique.
authors.affiliation: a Animal Biotechnology Center , Nanaji Deshmukh Veterinary Science University , Jabalpur , Madhya Pradesh , India.
authors.forename: Jyoti Lakshmi
authors.initials: JL
authors.lastname: Hati Boruah
authors.affiliation: a Animal Biotechnology Center , Nanaji Deshmukh Veterinary Science University , Jabalpur , Madhya Pradesh , India.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Ranjan
authors.affiliation: a Animal Biotechnology Center , Nanaji Deshmukh Veterinary Science University , Jabalpur , Madhya Pradesh , India.
authors.forename: Hamen
authors.initials: H
authors.lastname: Gogoi
authors.affiliation: a Animal Biotechnology Center , Nanaji Deshmukh Veterinary Science University , Jabalpur , Madhya Pradesh , India.
authors.forename: Saurabh Kumar
authors.initials: SK
authors.lastname: Pandey
authors.affiliation: a Animal Biotechnology Center , Nanaji Deshmukh Veterinary Science University , Jabalpur , Madhya Pradesh , India.
authors.forename: Dharmendra
authors.initials: D
authors.lastname: Kumar
authors.affiliation: a Animal Biotechnology Center , Nanaji Deshmukh Veterinary Science University , Jabalpur , Madhya Pradesh , India.
authors.forename: Amlan Jyoti
authors.initials: AJ
authors.lastname: Phukan
authors.affiliation: a Animal Biotechnology Center , Nanaji Deshmukh Veterinary Science University , Jabalpur , Madhya Pradesh , India.
authors.forename: Joygeswar
authors.initials: J
authors.lastname: Bori
authors.affiliation: a Animal Biotechnology Center , Nanaji Deshmukh Veterinary Science University , Jabalpur , Madhya Pradesh , India.
authors.forename: Bikash Chandra
authors.initials: BC
authors.lastname: Sarkhel
mesh_terms: D000818:Animals; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000235:genetics; D053148:Caspase 3 / Q000235:genetics; D002478:Cells, Cultured; D005333:Fetus / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D055785:Gene Knockdown Techniques / Q000379:methods*; D006041:Goats; D007372:Interferons / Q000378:metabolism; D055435:Myostatin / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000379:methods*
chemical_list: D051017:Apoptosis Regulatory Proteins; C525911:MSTN protein, human; D055435:Myostatin; D034741:RNA, Small Interfering; D007372:Interferons; D053148:Caspase 3
keywords: Apoptosis; RNA interference; caprine fetal fibroblasts; co-transfection; interferon; knockdown; myostatin; short hairpin RNA