Identification and functional characterization of long non-coding RNAs regulated by micro-RNAs

• Grant number: 13/14550-2
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: January 01, 2014
• End date: October 31, 2014
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Sergio Verjovski Almeida
• Grantee: Murilo Sena Amaral
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Recent works have shown that the major part of the messages transcribed in human cells is composed of non-protein coding RNAs (ncRNAs). A large fraction of these ncRNAs is composed of short (< 200 nucleotides) RNAs. Among these, the micro-RNAs (miRNAs) have been more extensively studied, as they are predicted to regulate over 60% of the protein-coding RNAs in humans. Another subclass of ncRNAs, named long (> 200 nucleotides) non-coding RNAs (lncRNAs), is transcribed from intergenic and mainly intronic regions of the human genome and has a regulatory function in normal and pathological conditions. Recently, the structure and function of a small set of lncRNAs have been characterized. However, the mechanisms by which the lncRNA levels are regulated are poorly understood. The hypothesis to be tested in this project is that lncRNAs are negatively regulated by miRNAs and that this regulation is direct and widespread along the human transcriptome. The experiments proposed initially aim to identify lncRNAs that are possible targets of miRNAs using high-throughput sequencing of lncRNAs bound to the protein argonaute2. Further, we intend to validate by gene-reporter assays and anti-miR assays the interaction between miRNAs and lncRNAs to be selected, and to confirm the direct regulation of lncRNAs by miRNAs. Finally, the functional role of the possible regulation miRNA-lncRNA will be tested in a cellular model using cell proliferation, cell cycle and apoptosis assays. Taken together, the experiments proposed aim to clarify the role of miRNAs in silencing the lncRNAs and in modulating their functional roles and, in a broader perspective, to contribute to the understanding of the metabolism and the regulation of the levels of lncRNAs in human cells. (AU)