Poster abstracts
Poster number 167 submitted by Kate Desrochers
Understanding the Modification Landscape of Anticodon-Engineered tRNAs and their Implications for Cystic Fibrosis Treatment
Kate Desrochers (Chemistry Department, University of Michigan), Bojing Zhu (Chemistry Department, University of Michigan), Kari Thrasher (Pharmacology and Physiology, University of Rochester), Daniel E. Eyler (Chemistry Department, University of Michigan), John Lueck (Pharmacology and Physiology, University of Rochester), Kristin S. Koutmou (Chemistry Department, University of Michigan)
Abstract:
Cystic Fibrosis (CF) is an autosomal recessive disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR). Premature termination codons (PTCs) lead to the synthesis of truncated, and usually nonfunctional, proteins. Approximately 10% of CF patients carry a PTC mutation, and no currently available therapeutics address these mutations. It is believed that rescuing 15-30% of CFTR channel function would have a meaningful impact on the disease state for patients with PTC mutations. Anticodon engineered tRNAs (ACE-tRNAs) are being developed to enable an amino acid to be incorporated at a PTC. The goal of this technology is to promote the synthesis of full-length CFTR protein from a mutated mRNA containing a PTC. While ACE-tRNAs have been shown to work robustly in vitro, the molecular determinants that guide the efficiency and efficacy of ACE-tRNAs are not fully understood. In particular, the contributions (if any) of post-transcriptional modifications are key for canonical tRNA stability, folding, transport and function, in ACE-tRNA function remain to be established. This project aims to elucidate the modification landscape of ACE-tRNAs expressed in mammalian cells through the use of Nanopore sequencing and mass spectrometry. This work will provide a better understanding of how the post-transcriptional status and availability of native and therapeutic tRNAs influence PTC readthrough. This information will fill critical gaps in the development of novel tRNA therapeutics for Cystic Fibrosis, as well as other genetic diseases caused by nonsense mutations.
Keywords: Cystic Fibrosis, tRNAs, RNA modifications