Poster abstracts
Poster number 130 submitted by Shashi Singh
Dual-Action Circular ADAR-Recruiting RNAs: A Novel Approach for Reversible Gene Modulation through RNA Editing and Antisense Effects
Shashi S. Singh (Department of Biochemistry and Center for RNA Science and Therapeutics, Case Western Reserve University, Cleveland, OH), Joseph M. Luna (Department of Biochemistry and Center for RNA Science and Therapeutics, Case Western Reserve University, Cleveland, OH)
Abstract:
RNA editing offers a promising approach for reversible gene modulation, addressing the limitations of permanent genome editing. We present a novel strategy using circular ADAR-recruiting guide RNAs (circ-arRNAs) to harness endogenous Adenosine Deaminase Acting on RNA (ADAR) activity for precise adenosine-to-inosine editing. These self-cleaving, self-ligating circ-arRNAs bind specific transcripts, recruiting ADAR to induce targeted base edits. We demonstrate circ-arRNA efficacy in two key applications: (1) stop codon removal in a fluorescent reporter system, and (2) exon skipping in KRAS through intron-exon boundary editing. Direct RNA delivery of circ-arRNAs showed superior editing kinetics compared to plasmid-based methods. Unexpectedly, we observed antisense oligonucleotide (ASO)-like activity of circ-arRNAs, with splice site switching occurring even in ADAR knockout cells, indicating a mechanism independent of RNA editing. This dual functionality of circ-arRNAs - combining ADAR-mediated editing and ASO-like effects - presents a versatile tool for manipulating gene expression. We are developing a bichromatic reporter system to assess circ-arRNA-mediated effects in various cancer cell lines. This approach holds significant potential for therapeutic applications, particularly in scenarios where transient modulation of gene expression is preferable to permanent genetic alterations.
Keywords: RNA Editing, Circular RNAs, Splicing modifier