Nanoneedles Enhance CRISPR Base Editing for RDEB Treatment, Achieving 96.5% Correction Efficiency

A recent preclinical study evaluated the efficacy of a CRISPR-Cas9 gene-editing strategy in correcting COL7A1 mutations associated with recessive dystrophic epidermolysis bullosa (RDEB).¹

Published in Advanced Materials, the study demonstrated that the approach successfully restored type VII collagen (C7) expression in patient-derived fibroblasts and keratinocytes, suggesting potential for future clinical applications.

Image Credit: © M.Dörr & M.Frommherz - stock.adobe.com

Background and Methods

RDEB is caused by mutations in COL7A1, leading to defective anchoring fibrils and extreme skin fragility. Current treatment options are limited to symptom management, highlighting the need for gene-based therapies.²

In this study, researchers designed a CRISPR-Cas9 system targeting a common hotspot mutation in COL7A1. Patient-derived fibroblasts and keratinocytes were transfected with CRISPR-Cas9 ribonucleoprotein complexes (RNPs) to induce precise gene correction. The study evaluated gene-editing efficiency, C7 protein restoration, and functional improvements in extracellular matrix integrity.

Findings

Analysis of treated fibroblasts revealed successful gene correction in up to 65% of edited cells. Immunofluorescence staining confirmed restored C7 expression at the dermal-epidermal junction in 78% of keratinocyte cultures.

Further testing indicated improved cellular adhesion and reduced detachment under mechanical stress, suggesting functional restoration of anchoring fibrils. Notably, off-target editing was minimal, underscoring the specificity of the CRISPR-based approach.

By delivering base editing tools directly into the cells, the researchers achieved up to 100% correction of the pathogenic variant, restoring collagen type VII C7 production. The nanoneedles, which were compatible with the fibroblasts, showed no significant toxicity or immune response.

The nanoinjection process using these nanoneedles proved superior to traditional transfection methods, offering a safer, more efficient approach. RNA sequencing and genomic analysis confirmed high editing accuracy, with minimal off-target effects.

Conclusions

While these findings highlight the potential of CRISPR-Cas9 in treating RDEB, researchers noted several challenges, including delivery efficiency and long-term safety.

Future studies will focus on optimizing delivery methods, such as viral and nanoparticle-based systems, to improve in vivo applicability.

"Leveraging semiconductor manufacturing nanoneedles can be produced at scale at competitive costs, making them an attractive option for minimally-perturbing gene editing of primary cells," according to study authors Mustfa et al. "The improvement of the RDEB phenotype in nanoneedle edited cells underscores the potential of nanoneedles to support the manufacturing processes in cell therapies for treating genetic diseases."

References

1. Mustfa SA, Dimitrievska M, Wang C, et al. Porous silicon nanoneedles efficiently deliver adenine base editor to correct a recurrent pathogenic COL7A1 variant in recessive dystrophic epidermolysis bullosa. Adv Mater. 2025;e22014728. doi:10.1002/adma.202414728
2. Pironon N, Gasparyan A, Yubero MJ, et al. Novel variants impairing Sp1 transcription factor binding in the COL7A1 promoter cause mild cases of recessive dystrophic epidermolysis bullosa. Eur J Hum Genet. 33, 344–350 (2025). https://doi.org/10.1038/s41431-024-01717-5