RNA Interference: For Improving Traits and Disease Management in Plants

Kumar, Sanjeev; Dey, Avishek; Yau, Yuan‐Yeu; Easterling, Mona; Sahoo, Lingaraj

doi:10.1007/978-981-15-5228-1_14

Cited by 2 publications

(1 citation statement)

References 133 publications

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“…RNA interference (RNAi) has become one of the powerful tools in recent times to suppress the expression of speci c gene of interest [17][18], as a therapeutic option in disease management [19][20][21][22], as cancer treatment [23][24][25] and for understanding regulatory function of a gene by dsRNA molecules [26]. Suppression of gene expression achieved by cleavage of dsRNA molecules by ribonuclease protein (Dicer) followed by loading of siRNA molecules into RISC (RNA induced silencing complex) and then, guide strand of the siRNA molecule would guide the RISC towards the target mRNA sequence complementary with the siRNA for cleavage of target mRNA [27][28][29][30][31].…”

Section: Discussionmentioning

confidence: 99%

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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