Preprint
Preventing vision loss in a mouse model of Leber Congenital Amaurosis by engineered tRNA
bioRxiv
07/11/2025
DOI: 10.1101/2025.07.10.660754
PMCID: PMC12265521
PMID: 40672310
Abstract
Premature termination codons (PTCs) are associated with rare genetic disorders. Inducing targeted read-through of these 'nonsense mutations' presents a potential therapeutic strategy for modifying disease outcomes. We previously reported that one such PTC, W53X, in the KCNJ13 gene causes blindness and Leber congenital amaurosis type-16 (LCA-16) due to loss of function of the inwardly rectifying potassium channel 7.1 (Kir7.1). Here, we present the proof of concept of a therapeutic approach based on anticodon-engineered transfer RNA (ACE-tRNA). The ACE-tRNA encodes the amino acid tryptophan (Trp) and suppresses the W53X PTC, restoring full-length protein expression. We used helper-dependent adenovirus (HDAd) to deliver the ACE-tRNA Trp.UAG (tRNA Trp.UAG ) and rescue Kir7.1 function and physiology in patient-specific human induced pluripotent stem cell-derived retinal pigment epithelium (hiPSC-RPE) cells. Furthermore, in a W53X mouse model of LCA16, HDAd delivery of tRNA Trp.UAG resulted in durable restoration of vision as measured by retinography. This study provides the first example of the therapeutic application of ACE-tRNA for treating an inherited form of blindness.Premature termination codons (PTCs) are associated with rare genetic disorders. Inducing targeted read-through of these 'nonsense mutations' presents a potential therapeutic strategy for modifying disease outcomes. We previously reported that one such PTC, W53X, in the KCNJ13 gene causes blindness and Leber congenital amaurosis type-16 (LCA-16) due to loss of function of the inwardly rectifying potassium channel 7.1 (Kir7.1). Here, we present the proof of concept of a therapeutic approach based on anticodon-engineered transfer RNA (ACE-tRNA). The ACE-tRNA encodes the amino acid tryptophan (Trp) and suppresses the W53X PTC, restoring full-length protein expression. We used helper-dependent adenovirus (HDAd) to deliver the ACE-tRNA Trp.UAG (tRNA Trp.UAG ) and rescue Kir7.1 function and physiology in patient-specific human induced pluripotent stem cell-derived retinal pigment epithelium (hiPSC-RPE) cells. Furthermore, in a W53X mouse model of LCA16, HDAd delivery of tRNA Trp.UAG resulted in durable restoration of vision as measured by retinography. This study provides the first example of the therapeutic application of ACE-tRNA for treating an inherited form of blindness.
Details
- Title: Subtitle
- Preventing vision loss in a mouse model of Leber Congenital Amaurosis by engineered tRNA
- Creators
- Enes Akyuz - University of Wisconsin–MadisonPawan K Shahi - University of Wisconsin–Fox ValleyLionel Gissot - University of IowaAhmad Al Saneh - University of IowaDivya Sinha - University of Wisconsin–MadisonGiovanni M Hanstad - University of Wisconsin–MadisonMeha Kabra - University of Wisconsin–MadisonMaria A Fernandez Zepeda - University of Wisconsin–MadisonDavid 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
- Preprint
- Publication Details
- bioRxiv
- DOI
- 10.1101/2025.07.10.660754
- PMID
- 40672310
- PMCID
- PMC12265521
- NLM abbreviation
- bioRxiv
- ISSN
- 2692-8205
- eISSN
- 2692-8205
- Language
- English
- Date posted
- 07/11/2025
- Academic Unit
- Molecular Physiology and Biophysics; Iowa Neuroscience Institute; Otolaryngology
- Record Identifier
- 9984847993302771
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