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3′UTR and promoter luciferase reporter assays

DC Dongdong Chen
JC Julie Cox
JA Jayabhargav Annam
MW Melanie Weingart
GE Grace Essien
KR Komal S. Rathi
JR Jo Lynne Rokita
PK Priya Khurana
SC Selma M. Cuya
KB Kristopher R. Bosse
AP Adeiye Pilgrim
DL Daisy Li
CS Cara Shields
OL Oskar Laur
JM John M. Maris
RS Robert W. Schnepp
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Assays were carried out with the Lightswitch Luciferase Assay System (Switchgear Genomics) as previously detailed. We utilized control and let-7i microRNA mimetics (Dharmacon; listed in Supplementary Table S1) and the effects of control and let-7i microRNA mimetics on the PBK 3′UTR were normalized to effects on actin (which does not contain let-7 binding sites) [11]. PBK promoter luciferase assays were carried out as previously described, with PBK promoter luciferase values normalized to CellTiter-Glo luminescence assay values [18].

Copyright and License information:  ©2020 The Authors
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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