Short Hairpin RNA-Induced Myostatin Gene Silencing in Caprine Myoblast Cells In Vitro

Tripathi, Ajai K.; Ramani, Umed; Patel, Amrutlal K.; Rank, D. N.; Joshi, Chaitanya G.

doi:10.1007/s12010-012-0021-1

Cited by 17 publications

(11 citation statements)

References 19 publications

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“…According to McGivney et al (2012) and Tripathi et al (2013), the muscle growth process unites hyperplasia and hypertrophy, and is regulated by myogenic factors. Myostatin is a member of the genes of the transforming growth factor beta (TGF-β).…”

Section: Discussionmentioning

confidence: 99%

Effect of propolis ethanol extract on myostatin gene expression and muscle morphometry of Nile tilapia in net cages

Buck¹,

Mizubuti

Alfieri

et al. 2017

Genet. Mol. Res.

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ABSTRACT. Propolis can be used as growth enhancer due to its antimicrobial, antioxidant, and immune-stimulant properties, but its effects on morphometry and muscle gene expression are largely unknown. The present study evaluates the influence of propolis on muscle morphometry and myostatin gene expression in Nile tilapia (Oreochromis niloticus) bred in net cages. Reversed males (GIFT strain) with an initial weight of 170 ± 25 g were distributed in a (2 x 4) factorial 2 E.L. Buck et al.Genetics and Molecular Research 16 (1): gmr16019404 scheme, with two diets (DPRO, commercial diet with 4% propolis ethanol extract and DCON, commercial diet without propolis, control) and four assessment periods (0, 35, 70, and 105 experimental days). Muscles were evaluated at each assessment period. Histomorphometric analysis classified the fiber diameters into four groups: <20 mm; 20-30 mm; 30-50 mm; and > 50 mm. RT-qPCR was performed to assess myostatin gene expression. Fibers < 20 µm diameter were more frequent in DPRO than in DCON at all times. Fiber percentages >30 µm (30-50 and > 50 µm) at 70 days were 25.39% and 40.07% for DPRO and DCON, respectively. There was greater myostatin gene expression at 105 days, averaging 1.93 and 1.89 for DCON and DPRO, respectively, with no significant difference in any of the analyzed periods. Propolis ethanol extract did not affect the diameter of muscle fibers or the gene expression of myostatin. Future studies should describe the mechanisms of natural products' effects on muscle growth and development since these factors are highly relevant for fish production performance.

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Section: Discussionmentioning

confidence: 99%

Effect of propolis ethanol extract on myostatin gene expression and muscle morphometry of Nile tilapia in net cages

Buck¹,

Mizubuti

Alfieri

et al. 2017

Genet. Mol. Res.

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“…Our observations indicated that the four MEF2 genes in the ricefield eel at earlier development (sizes: 5–45 g) were all upregulated; these perhaps implied that they had cooperative action, which is according to that reported by Potthoff and Olson (). This could indicate that myoblast proliferation and differentiation during the initial growth phases could be inferred by the high expression levels of the MEF2 gene family (Hu et al ; Tripathi et al ). Thus, the data obtained further our understanding of molecular processes between muscle cell differentiation and growth and MEF2 gene family regulation function in fish, at least in the ricefield eel.…”

Section: Discussionmentioning

confidence: 99%

Molecular Characterization, Spatial–Temporal Expression Profiles, and Injury‐responsive Regulation of Myocyte‐specific Enhancer Factor 2 Gene Family in the Ricefield Eel, Monopterus albus

Chen

Cheng

et al. 2018

J World Aquaculture Soc

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The myocyte‐specific enhancer factor 2 (MEF2) gene family members are well‐established regulators in the control of muscle repair, regeneration, and development. In this study, we cloned the CDS sequences of the four MEF2 family members in the ricefield eel, and their coding sequences (CDS) lengths were 1449 bp, 1419 bp, 1431 bp, and 1608 bp, respectively. Sequence analysis demonstrated that three of them shared four conserved protein domains, including a MADS‐Box, inside SRF‐TF domains, adjacent conserved MEF2 domains, the HJURP‐C domains, while MEF2B have no HJURP‐C domains. Phylogenetic tree analyses indicated that the MEF2 family members were clustered together with those of fish species, and MEF2A, MEF2C, and MEF2D were closely clustered together and kept away from MEF2B. Spatial–temporal expression profiles displayed that the messenger RNA (mRNA) levels of MEF2 were upregulated with the developmental stage and were especially highly expressed significantly in skeletal muscles. After subcutaneous injection of bupivacaine hydrochloride, mRNA levels of MEF2 family and rapidly performed injury‐responsive regulation after damage that firstly increased, then decreased until normal levels were recovered, the same as the change in muscle fiber cells. In conclusion, the MEF2 gene family plays a central role in muscle repair, regeneration, and development in fish, at least in the ricefield eel.

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“…The transgenic goat having myostatin knocked down offers the potential to enhance the meat production. To achieve this objective, earlier, we and other group reported successful knockdown of MSTN in in vitro culture system using shRNA against MSTN gene through transient transfection assay . The current study aimed to examine whether a plasmid vector, expressing myostatin‐targeting shRNA, could be able to continuously suppress endogenous MSTN expression after integrating into the genome of GFF cells.…”

Section: Introductionmentioning

confidence: 99%

Stable suppression of myostatin gene expression in goat fetal fibroblast cells by lentiviral vector‐mediated RNAi

Patel

Joshi

2014

Biotechnology Progress

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Myostatin (MSTN) is a secreted growth factor that negatively regulates skeletal muscle mass, and therefore, strategies to block myostatin-signaling pathway have been extensively pursued to increase the muscle mass in livestock. Here, we report a lentiviral vector-based delivery of shRNA to disrupt myostatin expression into goat fetal fibroblasts (GFFs) that were commonly used as karyoplast donors in somatic-cell nuclear transfer (SCNT) studies. Sh-RNA positive cells were screened by puromycin selection. Using real-time polymerase chain reaction (PCR), we demonstrated efficient knockdown of endogenous myostatin mRNA with 64% down-regulation in sh2 shRNA-treated GFF cells compared to GFF cells treated by control lentivirus without shRNA. Moreover, we have also demonstrated both the induction of interferon response and the expression of genes regulating myogenesis in GFF cells. The results indicate that myostatin-targeting siRNA produced endogenously could efficiently down-regulate myostatin expression. Therefore, targeted knockdown of the MSTN gene using lentivirus-mediated shRNA transgenics would facilitate customized cell engineering, allowing potential use in the establishment of stable cell lines to produce genetically engineered animals.

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Short Hairpin RNA-Induced Myostatin Gene Silencing in Caprine Myoblast Cells In Vitro

Cited by 17 publications

References 19 publications

Effect of propolis ethanol extract on myostatin gene expression and muscle morphometry of Nile tilapia in net cages

Effect of propolis ethanol extract on myostatin gene expression and muscle morphometry of Nile tilapia in net cages

Molecular Characterization, Spatial–Temporal Expression Profiles, and Injury‐responsive Regulation of Myocyte‐specific Enhancer Factor 2 Gene Family in the Ricefield Eel, Monopterus albus

Stable suppression of myostatin gene expression in goat fetal fibroblast cells by lentiviral vector‐mediated RNAi

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