Journal article
Engineered tRNA reduces vision loss in a mouse model of Leber congenital amaurosis
Signal transduction and targeted therapy, Vol.11(1), 225
06/09/2026
DOI: 10.1038/s41392-026-02793-3
PMCID: PMC13250130
PMID: 42265079
Abstract
Premature termination codons (PTCs) are a major class of pathogenic variants that underlie rare inherited disorders, including forms of childhood blindness. Therapeutic suppression of these “nonsense mutations” offers a gene- and position-agnostic strategy to restore protein function. Our previous work established that the W53X PTC in the KCNJ13 gene causes Leber congenital amaurosis type 16 (LCA16) by disrupting the inwardly rectifying potassium channel Kir7.1, leading to retinal pigment epithelium (RPE) dysfunction. Here, we present a proof-of-concept approach using anticodon-engineered transfer RNA (ACE-tRNA) to promote targeted translational readthrough. We engineered a suppressor tRNA encoding tryptophan (ACE-tRNATrp.UAG) to selectively recognize the UAG stop codon at the W53X site, enabling incorporation of the correct amino acid and restoration of full-length Kir7.1 protein. Delivery of ACE-tRNA via helper-dependent adenovirus (HDAd) resulted in robust rescue of channel function in heterologous systems expressing mutant KCNJ13 and in patient-derived human induced pluripotent stem cell (hiPSC)-RPE cells. Functional recovery was confirmed by electrophysiological assays demonstrating restored inwardly rectifying currents and membrane potential. Importantly, subretinal delivery of HDAd-ACE-tRNATrp.UAG in a W53X mouse model led to partial restoration of RPE physiology, as measured by electroretinography, without evidence of retinal toxicity. Together, these findings establish ACE-tRNA-mediated suppression as a viable therapeutic strategy for nonsense mutations in multimeric ion channels and provide a translational framework for precision treatment of inherited retinal diseases.
Details
- Title: Subtitle
- Engineered tRNA reduces vision loss in a mouse model of Leber congenital amaurosis
- Creators
- Pawan K. Shahi - University of Wisconsin–MadisonEnes Akyuz - University of Wisconsin–MadisonLionel Gissot - University of IowaAhmad Al Saneh - University of IowaSanjai K. Pillala - University of Wisconsin–MadisonDivya Sinha - University of Wisconsin–MadisonGiovanni M. Hanstad - University of Wisconsin–MadisonMeha Kabra - University of Wisconsin–MadisonMaria A. Fernandez Zepeda - Madison, WI USADavid M. Gamm - University of Wisconsin–MadisonSamuel M. Young - University of North Carolina at Chapel HillChristopher A. Ahern - University of IowaBikash R. Pattnaik - University of Wisconsin–Madison
- Resource Type
- Journal article
- Publication Details
- Signal transduction and targeted therapy, Vol.11(1), 225
- DOI
- 10.1038/s41392-026-02793-3
- PMID
- 42265079
- PMCID
- PMC13250130
- NLM abbreviation
- Signal Transduct Target Ther
- ISSN
- 2095-9907
- eISSN
- 2059-3635
- Publisher
- Nature Publishing Group
- Grant note
- U.S. Department of Health & Human Services | NIH | National Eye Institute (NEI): R24 EY032434
We used BioRender, licensed to UW-Madison and NIH BioArt, to generate the artwork shown in all the figures. This work was supported by the National Institute of Health (grant number R24EY032434), R01DC014093 (S.M.Y.), and R01NS110742 (S.M.Y.). This study was supported in part by the Retina Research Foundation, M.D. Matthews Professorship (B.R.P.), Daniel M. Albert Chair in Eye Research (B.R.P.), Sandra Lemke Trout Chair in Eye Research (D.M.G.), RRF Emmett A. Humble Distinguished Directorship (D.M.G.) of the McPherson Eye Institute, UW-Madison, Action for A-T, Million Dollar Bike Ride-Penn Medicine Orphan Disease Center, and UNC-CH start-up funds to S.M.Y. It was also partially supported by an Unrestricted Grant from Research to Prevent Blindness, Inc. to the UW-Madison Department of Ophthalmology and Visual Sciences. The authors also thank the National Institutes of Health (NIH) and the University of Wisconsin-Madison (P30 EY016665 and S10OD026957).
- Language
- English
- Date published
- 06/09/2026
- Academic Unit
- Molecular Physiology and Biophysics; Iowa Neuroscience Institute
- Record Identifier
- 9985174807302771
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