Talk abstracts

Talk on Friday 02:30-02:50pm submitted by Zhongxin Lu

A lentiviral-based genetic library for microRNA and its applications

Zhongxin Lu (Department of Biochemistry and Molecular Biology), Yanglong Zhu (Department of Biochemistry and Molecular Biology), Yong Li (Department of Biochemistry and Molecular Biology)

Abstract:
MicroRNAs (miRNA) are short 20-25 nucleotide RNA molecules that negatively regulate gene expression in animals and plants. Growing evidence indicates that microRNAs (miRNAs) are key participants in the cellular signaling pathway regulatory network. We have constructed the first lentiviral-based microRNA genetic library that consists of most human miRNA minigenes. Our modification of a third-generation Feline Immunodeficiency Virus (FIV)-based vector provides two genetic markers, neomycin and GFP, allowing both transient observation and stable selection. The lentiviral vector will also allow one to use packaged pseudoviral particles to transduce hard-to-transfect cells. We have utilized this genetic library to study NF-kB signaling pathways using a dual-reporter assay and identified a large number of microRNAs modulating NF-kB signaling. Using Western blotting and other analyses, we determined some microRNA targets in the canonical NF-kB signaling pathway. The present work reveals that microRNAs mediate critical signaling pathways, demonstrating added complexity of human signaling networks.

Keywords: microRNA, NF-kB signaling pathway, lentiviral-based genetic library