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 Skach; Paul B. McCray; Christopher A. Ahern

doi:10.1038/s41467-019-08329-4

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 Skach, Paul B. McCray, Christopher A. Ahern

Research output: Contribution to journal › Article

11 Scopus citations

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. Here we present a high-throughput, cell-based assay to identify anticodon engineered transfer RNAs (ACE-tRNA) which can effectively suppress in-frame PTCs and faithfully encode their cognate amino acid. In total, we identify 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 cystic fibrosis transmembrane conductance regulator (CFTR).

Original language	English (US)
Article number	822
Journal	Nature communications
Volume	10
Issue number	1
DOIs	https://doi.org/10.1038/s41467-019-08329-4
State	Published - Dec 1 2019
Externally published	Yes

ASJC Scopus subject areas

Chemistry(all)
Biochemistry, Genetics and Molecular Biology(all)
Physics and Astronomy(all)

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Lueck, J. D., Yoon, J. S., Perales-Puchalt, A., Mackey, A. L., Infield, D. T., Behlke, M. A., Pope, M. R., Weiner, D. B., Skach, W., McCray, P. B., & Ahern, C. A. (2019). Engineered transfer RNAs for suppression of premature termination codons. Nature communications, 10(1), [822]. https://doi.org/10.1038/s41467-019-08329-4

Engineered transfer RNAs for suppression of premature termination codons. / Lueck, John D.; Yoon, Jae Seok; Perales-Puchalt, Alfredo; Mackey, Adam L.; Infield, Daniel T.; Behlke, Mark A.; Pope, Marshall R.; Weiner, David B.; Skach, William; McCray, Paul B.; Ahern, Christopher A.

In: Nature communications, Vol. 10, No. 1, 822, 01.12.2019.

Research output: Contribution to journal › Article

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