RT Journal Article
SR Electronic
T1 Engineered transfer RNAs for suppression of premature termination codons
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 400127
DO 10.1101/400127
A1 John D. Lueck
A1 Jae Seok Yoon
A1 Alfredo Perales-Puchalt
A1 Adam L. Mackey
A1 Daniel T. Infield
A1 Mark A. Behlke
A1 Marshall R. Pope
A1 David B. Weiner
A1 William R. Skach
A1 Paul B. McCray, Jr.
A1 Christopher A. Ahern
YR 2018
UL http://biorxiv.org/content/early/2018/08/27/400127.abstract
AB 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 anticodon engineered 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).