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SOCS1 luciferase reporter assay (pre-miR-155)

YT Yuquan Tong
YL Yeongju Lee
XL Xiaohui Liu
JC Jessica L. Childs-Disney
BS Blessy M. Suresh
RB Raphael I. Benhamou
CY Chunying Yang
WL Weimin Li
MC Matthew G. Costales
HH Hafeez S. Haniff
SS Sonja Sievers
DA Daniel Abegg
TW Tristan Wegner
TP Tiffany O. Paulisch
EL Elizabeth Lekah
MG Maison Grefe
GC Gogce Crynen
MM Montina Van Meter
TW Tenghui Wang
QG Quentin M. R. Gibaut
JC John L. Cleveland
AA Alexander Adibekian
FG Frank Glorius
HW Herbert Waldmann
MD Matthew D. Disney
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HEK293T cells in 60-mm-diameter dishes (around 80% confluency) were co-transfected with 4 µg of a plasmid encoding SOCS1 fused to luciferase (Genecopoeia, HmiT021399-MT06) and 1 µg of the plasmid expressing wild-type pre-miR-155 using jetPRIME (Polypuls, 101000027) according to the manufacturer’s recommended protocol. The cells were seeded to 96-well plates (7,000 cells per well) after transfection. After incubating for 12 h, the cells were treated with DMSO (vehicle, 0.1% (v/v)) or pre-miR-155-RIBOTAC. A dose–response was measured by treating pre-miR-155-RIBOTAC at 0, 10 and 100 nM for 48 h and a time course was measured by treating pre-miR-155-RIBOTAC (100 nM) for 6, 24, 48 and 72 h. The Dual-Glo Luciferase Assay System (Promega, E2920) was used to measure luciferase activity, according to the manufacturer’s instructions.

Copyright and License information: The Author(s) ©2023
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