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. 2024 Mar 19;16(4):805–822. doi: 10.1038/s44321-024-00053-x

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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Fam161a^tm1b/tm1b mice were subretinally injected with 10¹⁰ GC of FCBR2-F0.4-HS (A), FCBR1-F0.4-HS (B), or HL (C) vectors between PN15 and PN26 and FAM161A expression was analyzed 2–4 months later. In total, 10¹¹ GC were also tested for the FCBR1-F0.4-HS (D), and HL (E) vectors. Note for all conditions, the appearance of FAM161A-positive CC (white arrows), but the expression of the HS isoform induced ectopic expression of the protein (orange arrows). (F, G) The co-injection of FCBR1-F0.4-HS and FCBR1-F0.4-HL produced a targeted and homogenous expression of the FAM161A in CC (white arrows) with some expression in the OPL as previously described in WT (green arrow), but also sometimes stronger labeling (orange arrow). (H) The vector combination (HS + HL) rescued the best ONL thickness compared to GFP-treated animals (n = 16, P < 0.001). FCBR1-F0.4-HS (n = 5) or HL (n = 7) also significantly protected the ONL against degeneration (mean ± SEM, P ≤ 0.05, P < 0.01, respectively). One-way ANOVA Kruskal–Wallis test followed by Dunn’s test for multiple comparisons between the groups; *P < 0.05; **P < 0.01; ***P < 0.001. Scale bars are 25 μm. ONL outer nuclear layer, HS short isoform, HL human long isoform. white arrows: correct labeling; orange arrows: modified labeling. Data information: Results in (A–G) are representative of five (FCBR2-F0.4), and five (FCBR1-F0.4-HS 10¹⁰ GC), three (FCBR1-F0.4-HL 10¹⁰), three (FCBR1-F0.4-HS 10¹¹), four (FCBR1-F0.4-HL 10¹¹), and three (FCBR1-F0.4-HS + HL 10¹¹), independent experiments, n = 5 eyes to 16 per group as described in (H). Source data are available online for this figure.