Construction and Functional Analysis of lncRNAs RMST promoter luciferase reporter plasmid

Abstract

Long noncoding RNAs (lncRNAs) are abundant in the mammalian transcriptome and many are specifically expressed in the brain. Rhabdomyosarcoma 2-associated transcript (RMST), which are indispensable for neurogenesis. RMST expression is specific to the brain,regulated by the transcriptional repressor REST, and increases during neuronal differentiation, indicating a role in neurogenesis. RMST physically interacts with SOX2, a transcription factor known to regulate neural fate. RMST and SOX2 coregulate a large pool of downstream genes implicated in neurogenesis. It is known that the Myocardin can regulate the downstream gene transcription by combining the SRF-forming complex with the CArG box site of the target gene promoter. By analyzing the promoter sequence of RMST, we found that there were one CArG box binding sites withinthe promoter. In the present study, the gene fragment of human RMST gene promoter containing CArG box was cloned into empty vector pGL3-Basic to construct RMST promoter luciferase reporter plasmid. Myocardin expression plasmids, together with RMST promoter-luci plasmids were transfected into COS-7 cells, and then luciferase assay was performed to analyze the effects of Myocardin on regulating the promoter activity of RMST. The results showed that Myocardin can activate the transcription of RMST. Constuction of RMST promoter luciferase reporter plasmid will provide the theory basis for investigating the function of RMST in neurogenesis.