Porous Silicon Nanoneedles Efficiently Deliver Adenine Base Editor to Correct a Recurrent Pathogenic COL7A1 Variant in Recessive Dystrophic Epidermolysis Bullosa

Abstract

Base editing, a CRISPR-based genome editing technology, enables precise correction of single-nucleotide variants, promising resolutive treatment for monogenic genetic disorders like recessive dystrophic epidermolysis bullosa (RDEB). However, the application of base editors in cell manufacturing is hindered by inconsistent efficiency and high costs, contributed by suboptimal delivery methods. Nanoneedles have emerged as an effective delivery approach, enabling highly efficient, non-perturbing gene therapies both in vitro and in vivo. Here we demonstrate that nanoneedle delivery of an adenine base editor corrects a heterozygous single-nucleotide pathogenic variant in COL7A1 in primary RDEB fibroblasts in vitro with 96.5% efficiency, without inducing off-target variants. The nanoneedle delivery maintains cell viability and displays modest phenotypical alterations unlike conventional cationic lipid transfection. The nanoneedle-mediated editing significantly increases the production and secretion of full-length type VII collagen protein, contributing to restore functional fibroblasts phenotype by improving cell adhesion. These findings underscore the suitability and safety of nanoneedles for gene editing in a clinically relevant context of cell manufacturing, establishing a foundation for their use in cell therapies.