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Engineered transfer RNAs for suppression of premature termination codons
Preprint   Open access

Engineered transfer RNAs for suppression of premature termination codons

John D Lueck, Jae Seok Yoon, Alfredo Perales-Puchalt, Adam L Mackey, Daniel T Infield, Mark A Behlke, Marshall R Pope, David B Weiner, William R Skach, Paul B McCray Jr, …
bioRxiv: the preprint server for biology
Cold Spring Harbor Laboratory
08/27/2018
DOI: 10.1101/400127
url
https://doi.org/10.1101/400127View
Preprint (Author's original)This preprint has not been evaluated by subject experts through peer review. Preprints may undergo extensive changes and/or become peer-reviewed journal articles. Open Access

Abstract

Premature termination codons (PTCs) are responsible for 10-15% of all inherited disease. PTC suppression during translation offers a promising approach to treat a variety of genetic disorders, yet small molecules that promote PTC read-through have yielded mixed performance in clinical trials. We present a high-throughput, cell-based assay to identify a nti c odon e ngineered transfer RNAs (ACE-tRNA) which can effectively suppress in-frame PTCs and faithfully encode their cognate amino acid. In total, we identified ACE-tRNA with a high degree of suppression activity targeting the most common human disease-causing nonsense codons. Genome-wide transcriptome ribosome profiling of cells expressing ACE-tRNA at levels which repair PTC indicate that there are limited interactions with translation termination codons. These ACE-tRNAs display high suppression potency in mammalian cells, Xenopus oocytes and mice in vivo , producing PTC repair in multiple genes, including disease causing mutations within the cystic fibrosis transmembrane conductance regulator ( CFTR ).

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