Programmed repair of disease-causing UGA premature termination codons in mammalian brain

Ahmad Al Saneh; Lionel Gissot; Christopher A Ahern

doi:10.1093/nar/gkag695

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Programmed repair of disease-causing UGA premature termination codons in mammalian brain

Journal article

Open access

Peer reviewed

Programmed repair of disease-causing UGA premature termination codons in mammalian brain

Ahmad Al Saneh, Lionel Gissot and Christopher A Ahern

Nucleic acids research, Vol.54(13), gkag695

07/03/2026

DOI: 10.1093/nar/gkag695

PMID: 42460447

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Abstract

Protein-truncating variants caused by stop codons are the most prevalent class of rare variant mutations in neurodevelopmental diseases, with UGA codons being most common. Suppressor transfer RNA (sup-tRNA) has therapeutic potential for premature termination codon (PTC) rescue but has thus far underperformed by traditional AAV delivery platforms, and progress has been hampered by the lack of methods to non-invasively assess in vivo activity in mammalian brain. To fill this material gap, we utilize transcranial in vivo bioluminescence imaging data from a luciferase-UGA mouse model to optimize viral payloads with sup-tRNA genes. These data demonstrate that U6 promoter-driven and single-stranded AAV2/9 constructs show variable and dose-dependent activity, whereas self-complementary AAV2/9 with the tRNA in a minimal 100-bp genomic context provides broad and efficacious PTC rescue. Further, payload tRNA multiplexing and use of tRNA introns enable efficacy of low viral titers and sustained rescue. tRNA sequencing of scAAV-delivered ArgUCA sup-tRNA in brain demonstrates no effects on endogenous tRNA levels, their acylation, or processing, and these features are also maintained in the delivered ArgUCA sup-tRNA. Collectively, this work defines a scalable strategy for precision UGA PTC stop codon suppression, supporting development of durable genetic rescue therapies for neurodevelopmental disorders in the mammalian brain.

Animals

Brain - metabolism

Codon, Nonsense - genetics

Codon, Terminator

Dependovirus - genetics

Genetic Therapy - methods

Genetic Vectors - genetics

Humans

Mice

RNA, Transfer - genetics

RNA, Transfer - metabolism

Details

Title: Subtitle: Programmed repair of disease-causing UGA premature termination codons in mammalian brain
Creators: Ahmad Al Saneh - University of Iowa
Lionel Gissot - University of Iowa
Christopher A Ahern - University of Iowa
Resource Type: Journal article
Publication Details: Nucleic acids research, Vol.54(13), gkag695
DOI: 10.1093/nar/gkag695
PMID: 42460447
ISSN: 1362-4962
eISSN: 1362-4962
Publisher: Oxford University Press
Grant note: AR220030 / Simons Foundation 646844 / Simons Foundation Autism Research Initiative-Pilot Award P50HD103556 / Developmental Disabilities Research Center
Language: English
Date published: 07/03/2026
Academic Unit: Molecular Physiology and Biophysics; Iowa Neuroscience Institute
Record Identifier: 9985183130602771

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