The design, synthesis, and screening of ERK1/2-siRNA in rats
Xiao Xiong-shen
Abstract
Xiao Xiong-shen
Abstract
Objective To design and synthesize different kinds of small interfering RNA (siRNA) targeting different regions of extracellular signal-regulated kinase 1/2 (ERK1/2), and screen the siRNA that can specifically and effectively suppresse ERK1/2 expression. Methods Three ERK1 specific double stranded siRNAs and three ERK2 specific double stranded siRNAs were designed and transfected into vascular smooth muscle cells in rat by lipofectamine RNAiMAX reagent, the transcription level of ERK1/2 was analyzed by real-time fluorescence quantitative RT-PCR. Results The three ERK1 specific double stranded siRNAs and three ERK2 specific double stranded siRNAs could inhibit the expression of ERK1/2. The inhibiting rates of ERK1-siRNA1 and ERK2-siRNA2 in the lowest concentration group were the highest (95.77% and 93.23% , respectively). Conclusion ERK1/2-siRNAs was successfully designed and synthesized, which can specifically and effectively suppresse the expression of ERK1/2 gene.
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Objective To design and synthesize different kinds of small interfering RNA (siRNA) targeting different regions of extracellular signal-regulated kinase 1/2 (ERK1/2), and screen the siRNA that can specifically and effectively suppresse ERK1/2 expression. Methods Three ERK1 specific double stranded siRNAs and three ERK2 specific double stranded siRNAs were designed and transfected into vascular smooth muscle cells in rat by lipofectamine RNAiMAX reagent, the transcription level of ERK1/2 was analyzed by real-time fluorescence quantitative RT-PCR. Results The three ERK1 specific double stranded siRNAs and three ERK2 specific double stranded siRNAs could inhibit the expression of ERK1/2. The inhibiting rates of ERK1-siRNA1 and ERK2-siRNA2 in the lowest concentration group were the highest (95.77% and 93.23% , respectively). Conclusion ERK1/2-siRNAs was successfully designed and synthesized, which can specifically and effectively suppresse the expression of ERK1/2 gene.
Key concepts: Small interfering RNA, Lipofectamine, Transfection, Kinase, Molecular biology, RNA interference, Gene silencing, Cell biology