In silico prediction of short hairpin and in vitro silencing of activin receptor type IIB in chicken embryo fibroblasts by RNA interference
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Title |
In silico prediction of short hairpin and in vitro silencing of activin receptor type IIB in chicken embryo fibroblasts by RNA interference
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Creator |
Guru Vishnu, P., Bhattacharya, T.K., Bhushan, B., Kumar, P., Chatterjee, R.N., Paswan, C., Dushyanth, K., Divya, D. and Rajendra Prasad, A.
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Subject |
Muscle growth ACTRIIB Secondary structures Thermodynamic parameters Gene expression Gene silencing
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Description |
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Gene silencing by RNA interference is extensively used reverse genetic approach to analyse the implications of any gene in mammalian systems. The silencing of the Activin type IIB receptor belonging to transforming growth factor beta superfamily has demonstrated increase in muscle growth in many species. We designed five short hairpin RNA constructs targeting coding region of chicken ACTRIIB. All the shRNAs were transfected into chicken embryo fibroblast cells and evaluated their silencing efficiency by real time PCR and western blotting. Initially the computational analysis of target region and shRNA constructs was undertaken to predict sequence based features (secondary structures, GC% and H-b index) and thermodynamic features (ΔGoverall, ΔGduplex, ΔGbreak−target, ΔGintra−oligomer, ΔGinter−oligomer and ΔΔGends). We determined that all these predicted features were associated with shRNA efficacy. The invitro analysis of shRNA constructs exhibited significant (P Not Available |
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Date |
2019-12-05T04:59:56Z
2019-12-05T04:59:56Z 2019-06-01 |
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Type |
Research Paper
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Identifier |
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Not Available http://krishi.icar.gov.in/jspui/handle/123456789/27683 |
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Language |
English
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Relation |
Not Available;
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Publisher |
Not Available
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