Luciferase Reporter Assay

LINC00239 fragments containing wild-type (WT) or mutant (MUT) miR-484 binding sites were synthesized and cloned into the pmirGLO Dual-Luciferase Vector (Promega Corporation, Madison, WI, USA) to generate LINC00239-WT and LINC00239-MUT reporter vectors. Similarly, KLF12-WT and KLF12-MUT reporter vectors were obtained using the same experimental steps. For luciferase reporter assays, CRC cells were inoculated into 24-well plates and cotransfected with the WT or MUT reporter vector and miR-484 mimic or NC mimic using Lipofectamine^® 2000 transfection reagent. Forty-eight hours later, luciferase activity was determined with the Dual-Luciferase Reporter Assay System (Promega).