Targeted gene editing using non-viral methods could have significant advantages in terms of safe delivery of cargo and cost. Physical techniques to produce membrane disruption via nanoneedles have the capability to penetrate cell membranes and to deliver gene-editing nucleases. However, the challenges for these systems include inconsistency of membrane penetration and slow processing throughputs, which lead to low transfection efficiency and poor viability of cell products. Here, we present an ordered array silicon nanoneedles with full control of size, pitch, and tip sharpness fabricated by combining nanosphere lithography with isotropic dry etching of silicon. Gene-encapsulated supramolecular nanoparticles are then tethered on the nanoneedle surfaces via molecular recognition. By integrating the nanoneedles with microfluidic devices, we demonstrate rapid, efficient, and non-toxic in vitro gene-delivery to model cell lines. The versatile and scalable nanostructured platform proposed here represent an ideal system for the genetic engineering of a variety of cell types that will empower research focused on gene therapies and enable translation of these cellular therapy approaches.

Nanosubstrate-mediated intracellular delivery for high-throughput gene modification

255th National Meeting (2018)

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