Knockdown of the myostatin gene by RNA interference in caprine fibroblast cells

Jain, Hemlata; Singh, Sanjeev; Kadam, Mahesh M.; Sarkhel, B. C.

doi:10.1016/j.jbiotec.2009.10.017

Cited by 32 publications

(15 citation statements)

References 22 publications

(21 reference statements)

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“…Variation in silencing efficiency was observed among the different plasmids, despite their similar ranking and GC contents (five-star ranking; 43–52% GC). Similar variation in silencing efficiency (up to 92%) has been reported in a caprine fibroblast cell line [30], [33]. In recent years, myostatin silencing in fibroblasts has also been demonstrated in other species.…”

Section: Discussionsupporting

confidence: 80%

“…For this reason, some researchers have not detected changes in myostatin protein expression using an RNAi approach [32], [33], [38], [57]. In the present study, the monoclonal anti-EGFP antibody was selected to test myostatin protein expression indirectly in 293FT cells, via the tag-protein method [58], [59].…”

Section: Discussionmentioning

confidence: 96%

“…Furthermore, study of myostatin function has raised the possibility that its inhibition might be useful for increasing muscle mass in agricultural applications. Recently, several researchers have reported efficient RNAi-mediated silencing of myostatin using shRNA vectors in various species [33]–[36]. In particular, fibroblasts have been successfully used to study myostatin expression [37]–[39].…”

Section: Discussionmentioning

confidence: 99%

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MicroRNA-Mediated Myostatin Silencing in Caprine Fetal Fibroblasts

et al. 2014

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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.

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Section: Discussionsupporting

confidence: 80%

Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

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MicroRNA-Mediated Myostatin Silencing in Caprine Fetal Fibroblasts

et al. 2014

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“…So many researchers make efforts to produce double-muscled livestock that have more muscle mass by developing methods to suppress the expression of myostatin gene through knockdown or RNA interference. Zhang et al (2007) and Jain et al (2010) have focused on using knockout methods to inhibit the myostatin gene in sheep using a recombinant myostatin. Knocking out this gene includes a large genomic deletion might affect the function of its contiguous genes.…”

Section: Effect Of the Number Of Myostatin Allele Copies Present In Amentioning

confidence: 99%

Correlation Analysis Between Myostatin Gene Polymorphisms and Carcass Traits in New Zealand Romney Sheep

Ibrahim

Hickford

2015

Egyptian Journal of Genetics and Cytology

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“…All PCR products were ligated into pMD18-T to construct shRNA expression vectors targeting EGFP or MSTN. siRNA target sequence for EGFP and MSTN was previously reported (Kim and Rossi, 2003;Jain et al 2010).…”

Section: Construction Of Shrna Plasmidsmentioning

confidence: 99%

Sheep 7SK promoter for short hairpin RNA expression

Ni¹,

Hu²,

Hazi³

et al. 2012

Electro Journal of Biotech

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Background: Gene silencing mediated by small interfering RNA (siRNA) has become a powerful biological tool for the regulation of gene expression. For the synthesis of siRNA by vector-based expression systems, several mammalian small nuclear RNA (snRNA) promoters have been cloned and shown different transcriptional efficiencies. Results: In this study, we identified a sheep 7SK snRNA (s7SK) promoter based on the highly conserved polymerase III promoter elements. Promoter activity was measured by promoter-driven shRNA expression to suppress expression of an exogenous reporter gene and endogenous sheep gene. Conclusions: The knock down assay demonstrated that the s7SK induced more stronger inhibition effect than human U6 and H1 promoters. The use of this native sheep 7SK promoter for shRNA expressionis an important component for development of RNAibased gene therapy and production of transgenic animals in sheep species.

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Knockdown of the myostatin gene by RNA interference in caprine fibroblast cells

Cited by 32 publications

References 22 publications

MicroRNA-Mediated Myostatin Silencing in Caprine Fetal Fibroblasts

MicroRNA-Mediated Myostatin Silencing in Caprine Fetal Fibroblasts

Correlation Analysis Between Myostatin Gene Polymorphisms and Carcass Traits in New Zealand Romney Sheep

Sheep 7SK promoter for short hairpin RNA expression

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