Direct delivery of Cas9 or base editor protein and guide RNA complex enables genome editing in the retina
Open accessAbstract
Genome editing by CRISPR-Cas holds promise for the treatment of retinal dystrophies . For therapeutic gene editing, transient delivery of CRISPR-Cas9 is preferable to viral delivery which leads to long-term expression with potential adverse consequences. Cas9 protein and its guide RNA , delivered as ribonucleoprotein (RNP) complexes, have been successfully delivered into the retinal pigment epithelium in vivo . However, the delivery into photoreceptors , the primary focus in retinal dystrophies , has not been achieved. Here, we investigate the feasibility of direct RNP delivery into photoreceptors and retinal pigment epithelium cells. We demonstrate that Cas9 or adenine-base editors complexed with guide RNA , can enter retinal cells without the addition of any carrier compounds. Once in the retinal cells, editing rates vary based on the efficacy of the guide RNA and the specific location edited within the genes. Cas9 RNP delivery at high concentrations, however, leads to outer retinal toxicity. This underscores the importance of improving delivery efficiency for potential therapeutic applications in the future.
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sgRNA-induced DSB in hiPS-RPE reached similar ratios to those observed in undifferentiated hiPSCs and exceeded efficiencies previously reported in the literature.
“The results described above demonstrated substantial sgRNA-induced DSB in hiPS-RPE, reaching similar ratios to those observed in undifferentiated hiPSCs and exceeding efficiencies previously reported in the literature [28].”
From Precise CRISPR/Cas9 and Cas12 Correction Using Lipoplexes in Retinal Models Derived from Patients with Inherited Retinal Dystrophies (2026) · cites this paper as [28]
The Results report sgRNA-mediated indels in mouse RPE, but do not report editing in hiPS-RPE or compare it with undifferentiated hiPSCs. The Discussion says the paper’s RPE editing rates were close to previous reports, not that they exceeded them. The indexed passages have no page numbers. (LLM, conf=0.82)
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Retinal organoids mirror CRISPR-Cas9 gene editing efficiency observed in vivo
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“The results described above demonstrated substantial sgRNA-induced DSB in hiPS-RPE, reaching similar ratios to those observed in undifferentiated hiPSCs and exceeding efficiencies previously reported in the literature [28].”
InfluentialÉdition génique par transfert de complexes ribonucléoprotéiques de CRISPR/Cas9 dans la rétine
Malki, H., Pulman, J., & Dalkara, D. (2025) · médecine/sciences · doi:10.1051/medsci/2025091
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- Authors
- Pulman, J., Botto, C., Malki, H., Ren, D., Oudin, P., De Cian, A., As, M., Izabelle, C., Saubamea, B., Forster, V., Fouquet, S., Robert, C., Portal, C., El-Amraoui, A., Fisson, S., Concordet, J.-P., & Dalkara, D.
- Affiliations
- Centre National de la Recherche Scientifique (FR)
- Inserm (FR)
- Corresponding author
- Pulman, Juliette
- ORCID
- Venue
- Molecular Therapy Nucleic Acids
- Publisher
- Elsevier BV
- Published
- 2024-12
- Volume
- 35
- Issue
- 4
- Pages
- 102349
- ISSN
- 2162-2531 (ISSN-L)
- PMC
- PMC11531619
- OpenAlex
- W4402969161
- Cited by
- 12
- References
- 49
- Citations by year
- 2026: 4 · 2025: 8
- FWCI
- 1.63
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Molecular Therapy — Nucleic Acids (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- HAL (submittedVersion, cc-by-nc-nd)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Île-de-France Regional Scientific Council
- French National Research Agency (ANR-10-LABX-65, ANR-18-IAHU-01)
- Foundation Fighting Blindness Inc (PPA-0922-0840-INSERM)
- AFM-Téléthon
- Foundation for Medical Research (FRM PLP201810007761, SPF201909009287)
- Sorbonne Université
- Foundation Fighting Blindness
- Association Française contre les Myopathies
- Institut National de la Santé et de la Recherche Médicale
- Fondation Voir et Entendre
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- CRISPR/Cas9, base editors, guide RNA, ribonucleoprotein delivery, retinal gene therapy, photoreceptors, retinal pigment epithelium, retinal dystrophy
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