title: Quantitative Evaluation of Myostatin Gene in Stably Transfected Caprine Fibroblast Cells by Anti-Myostatin shRNA.
abstract: Skeletal muscle is the major component of lean tissue that is used for consumption, and myostatin is a negative regulator of skeletal muscle growth. Downregulation of this gene therefore offers a strategy for developing superior animals with enhanced muscle growth. Knockdown of myostatin was achieved by RNA interference technology. The anti-myostatin shRNA were designed and stably transfected in caprine fibroblast cells. The reduced expression of target gene was achieved and measured in clonal fibroblast cells by real-time PCR. Two single-cell clones induced significant decrease of myostatin gene expression by 73.96 and 72.66 %, respectively (P < 0.05). To ensure the appropriate growth of transfected cell, seven media were tested. The best suited media was used for transfected fibroblast cell proliferation. The findings suggest that shRNA provides a novel potential tool for gene knockdown and these stably transfected cells can be used as the donor cells for animal cloning.
authors.affiliation: Animal Biotechnology Centre, Nanaji Deshmukh Veterinary Science University, JNKVV Campus, Krishi Nagar, Adhartal, Jabalpur, Madhya Pradesh, 482004, India, drsudhirjain@gmail.com.
authors.forename: Sudhir Kumar
authors.initials: SK
authors.lastname: Jain
authors.forename: Hemlata
authors.initials: H
authors.lastname: Jain
authors.forename: Dharmendra
authors.initials: D
authors.lastname: Kumar
authors.forename: Megha Kadam
authors.initials: MK
authors.lastname: Bedekar
authors.forename: Akhilesh Kumar
authors.initials: AK
authors.lastname: Pandey
authors.forename: Bikash Chandra
authors.initials: BC
authors.lastname: Sarkhel
mesh_terms: D000818:Animals; D001483:Base Sequence; D002452:Cell Count; D049109:Cell Proliferation; D002470:Cell Survival; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005786:Gene Expression Regulation; D020868:Gene Silencing; D006041:Goats / Q000235:genetics*; D008969:Molecular Sequence Data; D055435:Myostatin / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection
chemical_list: D055435:Myostatin; D034741:RNA, Small Interfering