Stem Cell Self-Renewal

lincRNA-ROR[LncRNA]

miR-145[miRNA]

OCT4[mRNA]

Embryonic Stem Cell

Homo sapiens (human)

Dev Cell 2013 Apr 15 25, 69-80 doi:10.1016/j.devcel.2013.03.002 PMID:23541921

Endogenous miRNA sponge lincRNA-ROR regulates Oct4, Nanog, and Sox2 in human embryonic stem cell self-renewal

Luciferase reporter assay

We demonstrate that a lincRNA, linc-RoR, may function as a key competing endogenous RNA to link the network of miRNAs and core TFs, e.g., Oct4, Sox2, and Nanog. We engineered linc-RoR-overexpressing plasmids that have either wild-type (WT) or mutant (Mut) transcripts with mutations in the two miR-145-binding sites. We found that the ectopically expressed linc-RoR WT reduced the concentration of miR-145 during the cotransfection of the linc-RoR expression vector and low concentration of miRNA mimics in HEK293 cells. We also found that linc-RoR expression levels were decreased after transfection of miR-145 mimics, which indicated that both miR-145 and linc-RoR were cleaved and degraded during this process. These data suggest that linc-RoR functions as an miRNA sponge to reduce the efficient concentration of miR-145. In addition, OCT4 and NANOG mRNA was rescued at up to 80% of the levels in wild-type hESCs after miR-145 inhibitor transfection.