Transfection of siRNA and CasRx Plasmid

Cells were transfected with the respective synthesized 21-mer siRNAs with dTdT or plasmids (Addgene Gene No. 237577 plasmid expressing CasRx and pSINmU6 encoding Grna) using the ScreenFect siRNA or ScreenFect A transfection kit (Fujifilm) in accordance with the manufacturer’s instructions, respectively [16 (link),31 (link)]. The medium was replaced 4 h after transfection and generally used for ≥48 h after transfection for biochemical experiments. In these conditions, attached cells incorporating trypan blue (Nacalai Tesque) were estimated to be <5% in each experiment 48 h after transfection [16 (link),31 (link)].

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Fukatsu S., Miyamoto Y., Oka Y., Ishibashi M., Shirai R., Ishida Y., Endo S., Katoh H, & Yamauchi J. (2023). Investigating the Protective Effects of a Citrus Flavonoid on the Retardation Morphogenesis of the Oligodendroglia-like Cell Line by Rnd2 Knockdown. Neurology International, 16(1), 33-61.

Publication 2023

ScreenFect A transfection kit trypan blue

Corresponding Organization : Tokyo Metropolitan Institute of Medical Science

Other organizations : Hayashibara (Japan), Tokyo Metropolitan University

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Variable analysis

independent variables

SiRNAs with dTdT
Plasmids (Addgene Gene No. 237577 plasmid expressing CasRx and pSINmU6 encoding Grna)

dependent variables

Biochemical experiments

control variables

Attached cells incorporating trypan blue (Nacalai Tesque) were estimated to be <5% in each experiment 48 h after transfection

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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