Gene Silencing Assay Using siRNAs

Mouse Smg1 and Smg7 siRNAs have been described previously^{5 (link)}. Mouse Eif2α (target sequence: 5′-CAAUGUUGUUAUGUUCU-3′) siRNA was designed using the i-Score Designer^{49 (link)} and asymmetric siRNA^{50 (link)} was synthesized (GeneDesign, Inc.). Other siRNAs, including mouse Atf4, FlexiTube siRNA (SI02674427), mouse Perk, FlexiTube siRNA (SI01319269), mouse Mtor, FlexiTube siRNA (SI03099796), and All Star Negative Control siRNA were purchased from Qiagen. Transfections of synthetic siRNAs were carried out with Lipofectamine RNAiMAX (Thermo Fisher Scientific) and the cells were analyzed 48 h after transfection. The results were confirmed in more than three independent experiments.

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Usuki F., Yamashita A, & Fujimura M. (2019). Environmental stresses suppress nonsense-mediated mRNA decay (NMD) and affect cells by stabilizing NMD-targeted gene expression. Scientific Reports, 9, 1279.

Publication 2019

FlexiTube siRNA Lipofectamine RNAiMAX

Atf4 Cells Lipofectamine Mouse Mtor Sirnas Transfection

Corresponding Organization : National Institute for Minamata Disease

Other organizations : Yokohama City University

Top 5 similar protocols

Variable analysis

independent variables

SiRNA knockdown of Smg1
SiRNA knockdown of Smg7
SiRNA knockdown of Eif2α
SiRNA knockdown of Atf4
SiRNA knockdown of Perk
SiRNA knockdown of Mtor

dependent variables

Measured outcomes after siRNA transfections (not explicitly specified)

control variables

Lipofectamine RNAiMAX transfection reagent
Timing of analysis (48 h after transfection)

controls

All Star Negative Control siRNA (negative control)

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