Abstract
CRISPR/Cas9-based gene-editing technologies offer promise for treating inherited retinal diseases (IRDs), however safe and efficient ocular delivery of precision editors remains challenging. To address this challenge, we report a class of Coomassie brilliant blue (CBB)-derived lipidoids that bind and deliver proteins. Subretinal injection of Cre complexed with these lipidoids into mT/mG mice leads to robust recombination in the retinal pigment epithelium and photoreceptors. We employ the CBB-lipidoid platform to deliver adenine base editor (ABE) ribonucleoproteins (RNP). Incorporating CBB lipidoids into liposomes improves delivery efficiency. CBB11 stands out for facilitating precise in vivo ABE-mediated gene editing. Delivery of liposome-CBB11-RNP complexes results in a 120-fold increase in base editing compared to RNP alone and restores the scotopic ERG b-wave response in the rd12 mouse model. These results demonstrate the potential of CBB-augmented, liposome-RNP systems for therapeutic gene editing in the eye, paving the way for single-dose precision medicines to treat IRDs.
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Data availability
The high-throughput sequencing data generated in this study were deposited in the National Center for Biotechnology Information Sequence Read Archive database under accession codes: PRJNA1240374 and PRJNA1358750. The data underlying the manuscript were deposited in Dryad: doi.org/10.5061/dryad.d51c5b0hd54 [https://datadryad.org/dataset/doi:10.5061/dryad.d51c5b0hd]. Source data are provided in this paper.
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Acknowledgements
We thank Marco Bassetto, Elliot H. Choi, Huajun B. Yan, and Alexander L. Yan for technical assistance. We acknowledge Dorota Skowronska-Krawczyk for access to a qPCR thermocycler, fluorescent microscope, and gel imager. We thank Suvrajit Sengupta at the UCI Department of Chemistry for technical support with NMR. We thank Jennifer Atwood and Michael Hou for technical assistance with flow cytometry. We thank Yekaterina Kadyshevskaya (University of Southern California) for the preparation of Fig. 1. We thank our colleagues at the UCI Center for Translational Vision Research and the Gavin Herbert Eye Institute for their comments regarding this manuscript. This work was supported in part by grants from the National Institutes of Health, including R01EY009339 (K.P.), 1R01EY034501 (K.P.), NSF-CHE-1904530 (G.P.T.), NIAID 75N93022C00054 (P.L.F.), DTRA grant N66001-21-C-4013 (P.L.F.), UG3AI150551 (D.R.L.), U01AI142756 (D.R.L.), R35GM118062 (D.R.L.), RM1HG009490 (D.R.L.), T32GM008620 (S.W.D.), F30EY033642 (S.W.D.); Foundation Fighting Blindness (award number TA-GT-0423-0847-UCI-TRAP) (K.P.); the Howard Hughes Medical Institute (HHMI) (D.R.L.); and Knights Templar Eye Foundation Career-Starter Research Grant (R.H.). R.H. is a Beckman Scholar in Retinal Research. The authors acknowledge support to the Department of Ophthalmology, Gavin Herbert Eye Institute at the University of California, Irvine, from an unrestricted Research to Prevent Blindness award, from NIH core grant P30EY034070, and from a University of California, Irvine School of Medicine Dean’s Office grant. The authors acknowledge support for the Chao Family Comprehensive Cancer Center’s Institute for Immunology Flow Cytometry Facility shared resource by the National Cancer Institute of the National Institutes of Health under award number P30CA062203. This article is subject to the Open Access to Publications policy of the Howard Hughes Medical Institute (HHMI). HHMI-supported authors have previously granted a nonexclusive CC BY 4.0 license to the public and a sublicensable license to HHMI in their research articles. Pursuant to those licenses, the author-accepted version of this manuscript can be made freely available under a CC BY 4.0 license immediately upon publication.
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Conceptualization was performed by J.Z., G.P.T, P.L.F., and K.P. Experimental investigations were performed by J.Z., S.W.D., J.H.F., R.S., R.H., G. P., G.P.T., C.R.M., Y.H., E.R., Z.D., X.M., M.H.S., L. X., M.H., P.Z.C., and B.L. Data analysis was performed by J.Z., R.H., S.W.D., J.H.F., R.S., C.R.M., E.R., G.P., and Y.H. Figures were prepared by J.Z., R.H., Y.H., G.P., and G.P.T. Manuscript was written by J.Z., R.H., Y.H., G.P.T., and K.P. Project was supervised by D.R.L., G.P.T., and K.P. Funds were acquired by R.H., S.W.D., D.R.L., G.P.T, P.L.F., and K.P. All authors contributed to the research and writing, and approved the manuscript.
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K.P. is a consultant for Polgenix Inc. and AbbVie Inc., and serves on the Scientific Advisory Board of Hyperion Eye Ltd. K.P. and G.P.T. are equity holders in Eyesomer Therapeutics. D.R.L. is a consultant and/or equity owner for Prime Medicine, Beam Therapeutics, Pairwise Plants, and nChroma Bio, companies that use or deliver genome-editing or epigenome-engineering agents. All other authors have declared that no competing interests exist.
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Zhang, J., Hołubowicz, R., Smidak, R. et al. A combinatorial synthetic strategy for developing genome-editing protein-delivery agents targeting mouse retina. Nat Commun (2026). https://doi.org/10.1038/s41467-026-69077-w
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DOI: https://doi.org/10.1038/s41467-026-69077-w
