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 journalArticle

2 Citations (Scopus)

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 languageEnglish (US)
Article number822
JournalNature communications
Volume10
Issue number1
DOIs
StatePublished - Dec 1 2019
Externally publishedYes

Fingerprint

ribonucleic acids
Nonsense Codon
Transfer RNA
Anticodon
retarding
cystic fibrosis
gametocytes
ribosomes
Repair
genome
Genes
regulators
Cells
mutations
genes
Cystic Fibrosis Transmembrane Conductance Regulator
mice
amino acids
disorders
Assays

ASJC Scopus subject areas

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

Cite this

Lueck, J. D., Yoon, J. S., Perales-Puchalt, A., Mackey, A. L., Infield, D. T., Behlke, M. A., ... 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 journalArticle

Lueck, JD, Yoon, JS, Perales-Puchalt, A, Mackey, AL, Infield, DT, Behlke, MA, Pope, MR, Weiner, DB, Skach, W, McCray, PB & Ahern, CA 2019, 'Engineered transfer RNAs for suppression of premature termination codons', Nature communications, vol. 10, no. 1, 822. https://doi.org/10.1038/s41467-019-08329-4
Lueck JD, Yoon JS, Perales-Puchalt A, Mackey AL, Infield DT, Behlke MA et al. Engineered transfer RNAs for suppression of premature termination codons. Nature communications. 2019 Dec 1;10(1). 822. https://doi.org/10.1038/s41467-019-08329-4
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. / Engineered transfer RNAs for suppression of premature termination codons. In: Nature communications. 2019 ; Vol. 10, No. 1.
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