Assessment of goat activin receptor type IIB knockdown by short hairpin RNAs<i>in vitro</i>

Patel, Amrutlal K.; Tripathi, Ajai K.; Shah, Rohan J.; Patel, Utsav; Joshi, Chaitanya G.

doi:10.3109/10799893.2014.922574

Cited by 4 publications

(2 citation statements)

References 35 publications

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“…In both the genes, the non-significant differences of expression of IFNA and IFNB genes between shRNA transfected cells and either scrambled shRNA transfected cells or un-transfected negative control embryonic hepatocyte cells. In supporting to the results obtained in the present study, Patel et al (2014) and Guru Vishnu et al (2019) in ACTIIB gene in goat and chicken fibroblast cells, respectively [26,51]. We suggest that shRNAs designed in our study against both SCD and SREBP1 genes had the potential to be excellent shRNA molecules for further development of knock down chicken as the shRNA molecules having very good knock down efficiency escaped the interferon system of the cells.…”

Section: Discussionsupporting

confidence: 91%

Prediction andin vitrosilencing of two lipid biosynthetic genes viz.SCDandSREBP1in chicken using RNA interference

Sagar

Prasad

Chatterjee

et al. 2020

Preprint

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RNA interference is a widely used post transcriptional silencing mechanism for suppressing expression of the target gene. In the current study, five shRNA molecules each against SCD and SREBP1 genes were designed after considering the parameters like secondary structures of shRNA constructs, mRNA target regions, GC content and thermodynamic properties (ΔG overall, ΔG duplex and ΔG break-target) to knockdown their expression. After successful transfection of these shRNA molecules into the chicken embryonic hepatocyte culture, expression of the target genes were monitored by real time PCR. Significant reduction (P<0.05) in the expression of SCD and SREBP1 genes with minimal activation of immune response genes in hepatocytes was observed after transfecting the shRNA molecules into them. The shRNA constructs against SCD gene showed the knock down efficiency ranged from 20.4% to 74.2%. In case of shRNA constructs against SREBP1 gene, they showed knock down efficiency ranging from 26.8% to 95.85%. These results clearly demonstrated the successful down regulation of the gene expression by designed shRNA molecules against both the target genes under in vitro condition. The shRNA2 molecule for SCD gene and the shRNA1 molecule for SREBP1 gene were found as the best among all the shRNA molecules used for silencing the target genes under cell culture system. It is concluded that the shRNA molecules designed against SCD and SREBP1 genes showed potential to silence the expression of these genes under in vitro chicken embryonic hepatocyte cell culture system.

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Section: Discussionsupporting

confidence: 91%

Prediction andin vitrosilencing of two lipid biosynthetic genes viz.SCDandSREBP1in chicken using RNA interference

Sagar

Prasad

Chatterjee

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…In both the genes, the non-signi cant differences of expression of IFNA and IFNB genes between shRNA transfecetd cells and either scrambled shRNA transfected cells or un-transfected negative control embryonic hepatocyte cells. In supporting to the results obtained in the present study, Patel et al (2014) and Guru Vishnu et al (2019) in ACTIIB gene in goat and chicken broblast cells, respectively [57,32]. We suggest that shRNAs designed in our study against both SCD and SREBP1 genes had the potential to be excellent shRNA molecules for further development of knock down chicken as the shRNA molecules having very good knock down e ciency escaped the interferon system of the cells.…”

Section: Discussionsupporting

confidence: 89%

S ilencing of SCD and SREBP1 Genes by Short Hairpin RNAs in Chicken Hepatic Cells in Vitro

Sagar

Prasad

Kumar

et al. 2022

Preprint

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RNA interference by short hairpin RNAs (shRNAs) is a widely used post transcriptional silencing mechanism for suppressing expression of the target gene. In the current study, five shRNA molecules each against SCD and SREBP1 genes involved in denovo lipid biosynthesis were designed upon considering parameters such as secondary structures of shRNAs, mRNA target regions, GC content and thermodynamic properties (ΔG overall, ΔG duplex and ΔG break-target), synthesized and cloned in pENTR/U6 entry vector to knockdown the expression of SCD and SREBP1 genes. After transfection of these shRNA constructs into the chicken embryonic hepatocytes, expressions of the target genes were monitored by real time PCR. Significant reduction (P<0.05) in the expression of SCD and SREBP1 genes was observed in hepatocytes. The shRNAs against SCD gene showed the knock down efficiency ranged from 20.4% (shRNA5) to 74.2% (shRNA2). In case of SREBP1 gene, the shRNAs showed knock-down efficiency ranging from 26.8% (shRNA4) to 95.85% (shRNA1). The shRNAs against both the genes introduced in chicken hepatocyte cells did not show any significant impact on expression of immune response genes (IFNA and IFNB) in those cells. These results clearly demonstrated the successful down regulation of the expression of SCD and SREBP1 genes by the shRNA molecules against both the target genes under in vitro condition. It is concluded that the shRNA molecules against SCD and SREBP1 genes showed great potential to silence the expression of these genes under in vitro chicken embryonic hepatocyte cells.

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In silico prediction of short hairpin RNA and in vitro silencing of activin receptor type IIB in chicken embryo fibroblasts by RNA interference

et al. 2019

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Assessment of goat activin receptor type IIB knockdown by short hairpin RNAsin vitro

Cited by 4 publications

References 35 publications

Prediction andin vitrosilencing of two lipid biosynthetic genes viz.SCDandSREBP1in chicken using RNA interference

Prediction andin vitrosilencing of two lipid biosynthetic genes viz.SCDandSREBP1in chicken using RNA interference

S ilencing of SCD and SREBP1 Genes by Short Hairpin RNAs in Chicken Hepatic Cells in Vitro

In silico prediction of short hairpin RNA and in vitro silencing of activin receptor type IIB in chicken embryo fibroblasts by RNA interference

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