Poster abstracts
Poster number 11 submitted by Daniel Dayeh
A programmable RNase for the targeted degradation of viral RNA
Daniel Dayeh (Chemistry & Biochemistry), William Cantara (Chemistry & Biochemistry), Karin Musier-Forsyth (Chemistry & Biochemistry), Kotaro Nakanishi (Chemistry & Biochemistry)
Abstract:
Since its discovery, RNA interference (RNAi) has enabled unprecedented advances to our understanding of cellular processes by employing small-interfering RNA (siRNA) to reduce endogenous protein levels for a gene product via degradation of messenger RNA (mRNA), yet its application towards other RNAs such as viral RNAs and long noncoding RNAs (lncRNAs) has been technically challenging. Here, we show that Argonaute (AGO), which plays a physiological role in RNAi in the budding yeast Kluyveromyces polysporus, can be programmed with a single-stranded guide DNA (gDNA) to site-specifically degrade the 5’ untranslated region of Human Immunodeficiency Virus - 1 (HIV-1 5’ UTR) by endonuclease activity. Site-specific degradation of viral RNAs will disrupt RNA:RNA and RNA:protein interactions by impairing or eliminating their binding sites, producing a map of the functional sites with nucleotide resolution. This approach is advantageous over existing methods due to the use of a DNA-induced slicing complex (DISC), which dramatically increases the stability of the DNA guides compared to the canonical RNA counterparts while reducing costs for largescale biotechnological applications. Overall, this strategy constitutes a high-throughput method for functional mapping of large, structured RNAs by identifying DISC-accessible sites, thus providing a platform for the investigation of physiologically and medically relevant RNAs.
Keywords: Argonaute