Poster abstracts

Poster number 25 submitted by Andrew Curtright

Elucidation of RNA functional motifs in a lncRNA by in vivo SELEX

Andrew J. Curtright (Department of Biochemistry, Case Western School of Medicine), Saba Valadkhan (Department of Biochemistry, Case Western School of Medicine)

Abstract:
Long non-coding RNAs (lncRNA) are a novel and abundant category of transcripts which are thought to play critical roles in regulation of diverse aspects of cellular function. Although a role for lncRNAs in several cellular processes has been described, the detailed mechanism behind the observed functions is still very poorly understood. The goal of this project is to advance our understanding of the sequence and structural elements which grant lncRNAs their specific functions. We have chosen a lncRNA as a study model for this project which we have shown to play a crucial role in stress response. In this project, we plan to introduce random mutations into the lncRNA using error-prone PCR. We will construct multiple pools of mutants with theoretical mutation rates which can vary from 50 to 1500 point mutations per molecule. Individual mutant constructs from this library will be cloned into lentiviral vectors and expressed in a pancreatic cell line and screened for resistance to oxidative stress. Mutants which confer a selective advantage against increased oxidative stress will be pooled, enriched and sequenced using high throughput sequencing technologies. The results obtained by this study will provide insights into the functional organization of the sequences within a lncRNA and the way non-coding RNAs can modulate this critical cellular pathway.

Keywords: SELEX, lncRNA, mutagenesis