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Cited by 20Retinal Development and DisordersCRISPR and Genetic EngineeringPhotoreceptor and optogenetics research

In vivo photoreceptor base editing ameliorates rhodopsin-E150K autosomal-recessive retinitis pigmentosa in mice

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Du, Samuel W., Newby, Gregory A., Salom, David, Gao, Fangyuan, Menezes, Carolline Rodrigues, Suh, Susie, Choi, Elliot H., Chen, Paul Z. +2 more·2024·Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.2416827121PMC PMC11621631
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Abstract

Rhodopsin, the prototypical class-A G-protein coupled receptor, is a highly sensitive receptor for light that enables phototransduction in rod photoreceptors. Rhodopsin plays not only a sensory role but also a structural role as a major component of the rod outer segment disc, comprising over 90% of the protein content of the disc membrane. Mutations in RHO which lead to structural or functional abnormalities, including the autosomal recessive E150K mutation, result in rod dysfunction and death. Therefore, correction of deleterious rhodopsin mutations could rescue inherited retinal degeneration, as demonstrated for other visual genes such as RPE65 and PDE6B. In this study, we describe a CRISPR/Cas9 adenine base editing strategy to correct the E150K mutation and demonstrate precise in vivo editing in a Rho -E150K mouse model of autosomal recessive retinitis pigmentosa (RP). Using ultraviolet-visible spectroscopy, mass spectrometry, and the G-protein activation assay, we characterized wild-type rhodopsin and rhodopsin variants containing bystander base edits. Subretinal injection of dual-adeno-associated viruses delivering our base editing strategy yielded up to 44% Rho correction in homozygous Rho -E150K mice. Injection at postnatal day 15, but not later time points, restored rhodopsin expression, partially rescued retinal function, and partially preserved retinal structure. These findings demonstrate that in vivo base editing can restore the function of mutated structural and functional proteins in animal models of disease, including rhodopsin-associated RP and suggest that the timing of gene-editing is a crucial determinant of successful treatment outcomes for degenerative genetic diseases.

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Cited by

Sorted by citations

Authors
Du, S. W., Newby, G. A., Salom, D., Gao, F., Menezes, C. R., Suh, S., Choi, E. H., Chen, P. Z., Liu, D. R., & Palczewski, K.
Affiliations
  • University of California, Irvine (US)
  • Broad Institute (US)
Corresponding author
  • Du, Samuel W.
  • Liu, David R.
  • Palczewski, Krzysztof
ORCID
Venue
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Published
2024-11-18
Volume
121
Issue
48
Pages
e2416827121-e2416827121
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
DOI
10.1073/pnas.2416827121
PMC
PMC11621631
OpenAlex
W4404469966
Cited by
20
References
9
Citations by year
2026: 11 · 2025: 9
FWCI
2.72
Type
journal article
Language
en
Open access
Licence
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • eScholarship (California Digital Library) (submittedVersion, cc-by)
Funders
HHS | NIH (R01EY009339 R01EY034501 P30EY034070, T32GM008620 F30EY033642, F30EY029136 T32GM007250 T32AR080622, R00HL163805, UG3AI150551 U01AI142756 R35GM118062 RM1HG009490)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Photoreceptor and optogenetics research
Keywords
rhodopsin, autosomal recessive retinitis pigmentosa, adenine base editing, CRISPR/Cas9, in vivo gene editing, retinal degeneration, rod photoreceptors, subretinal injection, G protein activation
MeSH terms
Gene Editing, Animals, Disease Models, Animal, Mutation, Retinitis Pigmentosa, Rhodopsin, Retinal Rod Photoreceptor Cells, Mice, CRISPR-Cas Systems

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