Dual luciferase reporting test

XX Xingcheng Xiong
JF Jiarui Feng
XY Xiao Yang
HL Hanjun Li
QS Qiao Shi
JT Jing Tao
JC Jian Chang
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By investigating the Encyclopedia of RNA Interactomes (http://starbase.sysu.edu.cn/) database, we obtained binding sites between miR-432-5p and circCDR1as or E2F3. The circular RNA CDR1as and E2F3 sequences containing the miR-432-5p binding sites were inserted into the luciferase reporter vectors (WT-circCDR1as, WT-E2F3). The site-specific mutation of the miR-432-5p binding sites in the circCDR1as and E2F3 sequences were also inserted into the luciferase reporter vectors (MUT-circCDR1as, MUT-E2F3). PANC1 and ASPC1 cells were co-transfected with miR-432-5p mimic or NC mimic with WT-circCDR1as/MUT-circCDR1as, WT-E2F3/MUT-E2F3 using Lipofectamine 2000. Finally, the luciferase activity was detected 24 h after transfection using Dual-Luciferase Reporter Assay System (Promega, Madison, WI, USA).

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