Journal article
Engineered transfer RNAs for suppression of premature termination codons
Nature communications, Vol.10(1), pp.822-822
02/18/2019
DOI: 10.1038/s41467-019-08329-4
PMCID: PMC6379413
PMID: 30778053
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).
Details
- Title: Subtitle
- Engineered transfer RNAs for suppression of premature termination codons
- Creators
- John D Lueck - Department of Physiology and Pharmacology, University of Rochester School of Medicine and Dentistry, Rochester, NY, 14642, USA. john_lueck@urmc.rochester.eduDaniel T Infield - Department of Molecular Physiology and Biophysics, Iowa Neuroscience Institute, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USAJae Seok Yoon - CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, 02421, MA, USAPaul B McCray Jr - Stead Family Department of Pediatrics, Pappajohn Biomedical Institute, University of Iowa, Iowa City, IA, 52242, USAAlfredo Perales-Puchalt - The Wistar Institute, Philadelphia, 19104, PA, USAChristopher A Ahern - Department of Molecular Physiology and Biophysics, Iowa Neuroscience Institute, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USA. christopher-ahern@uiowa.eduAdam L Mackey - Department of Molecular Physiology and Biophysics, Iowa Neuroscience Institute, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USAMark A Behlke - Integrated DNA Technologies Inc., Coralville, IA, 52241, USAMarshall R Pope - Department of Molecular Physiology and Biophysics, Iowa Neuroscience Institute, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USADavid B Weiner - The Wistar Institute, Philadelphia, 19104, PA, USAWilliam R Skach - Cystic Fibrosis Foundation, Bethesda, 20814, MD, USA
- Resource Type
- Journal article
- Publication Details
- Nature communications, Vol.10(1), pp.822-822
- DOI
- 10.1038/s41467-019-08329-4
- PMID
- 30778053
- PMCID
- PMC6379413
- NLM abbreviation
- Nat Commun
- ISSN
- 2041-1723
- eISSN
- 2041-1723
- Publisher
- England
- Grant note
- P30 CA010815 / NCI NIH HHS P30 DK054759 / NIDDK NIH HHS
- Language
- English
- Date published
- 02/18/2019
- Academic Unit
- Molecular Physiology and Biophysics; Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Medicine Administration; Internal Medicine
- Record Identifier
- 9984070706202771
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