Luciferase Assay of miR-545-3p Targets

The wide-type (WT) complementary sequence of FAM83H-AS1 or 3ʹUTR HS6ST2 to miR-545-3p and the matched mutant (MUT) sequence were amplified and ligated into pmirGLO vectors to produce FAM83H-AS1-WT, FAM83H-AS1-MUT, 3ʹUTR HS6ST2-WT and 3ʹUTR HS6ST2-MUT luciferase vectors. These constructed vectors were accompanied by miR-545-3p mimic or mimic NC to transfect HCC827 and NCI-H1650 cells. Finally, the relative luciferase activities were measured after 48 h of transfection using a Dual-Luciferase Assay Kit (Yeasen, China) [39 (link)].

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Zhang Y., Yu Y., Cao X, & Chen P. (2022). Role of lncRNA FAM83H antisense RNA1 (FAM83H-AS1) in the progression of non-small cell lung cancer by regulating the miR-545-3p/heparan sulfate 6-O-sulfotransferase (HS6ST2) axis. Bioengineered, 13(3), 6476-6489.

Publication 2022

Dual-Luciferase Assay Kit

3 utr Assay Cells Hs6st2 Luciferase Transfection Vectors

Corresponding Organization : Tianjin Medical University Cancer Institute and Hospital

Other organizations : Hebei North University

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Variable analysis

independent variables

MiR-545-3p mimic
Mimic NC (negative control)

dependent variables

Relative luciferase activities

control variables

FAM83H-AS1-WT
FAM83H-AS1-MUT
3'UTR HS6ST2-WT
3'UTR HS6ST2-MUT
HCC827 cells
NCI-H1650 cells

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