Gene delivery to animal and plant cells using a nano-needle array-shaped photooxidative cell membrane perforator

To avoid some cancer risks associated with the viral gene transfer process, the development of physical or chemical processes has received a great deal of attention in recent years. We found a unique cell membrane perforation method that applies photochemical oxidation to a limited cell membrane area, resulting in a high animal cell survival rate. We recently developed a nano-needle array cell membrane perforator that contains a photosensitizer for mass cell processing in a robotic system. In this research, we extended the gene transfer method to animal and plant cells. The targeted cells are human embryonic carcinoma cell line NT2/D1, onion epidermal cells, rice protoplasts and tobacco root calli. The cells were transformed with green fluorescent protein-expressing genes, and the transient expression of the genes was analyzed. Transient gene expression occurred and the adequate parameters for the expression were 400 gf/cm2 for 30 s for animal cells. The pressure and duration were in inverse proportions. The results in plant cells and protoplasts had similar parameters, but the plant cell walls required a relative high pressure, more than 50,000 gf/cm2.