Modulated substrate-controlled plasmid transfer to cell monolayer

Abstract

Efficient and controlled gene delivery remains a critical requirement in biomedical research, particularly for applications in tissue engineering, gene therapy and cell-based assays. Conventional non-viral gene delivery methods often face limitations such as low transfection efficiency, lack of spatial control and cytotoxicity. Hence there is need for innovative platforms that enable precise, efficient and localized gene transfer. In this study, in-house developed Poly(N-isopropylacrylamide)-Co-Glycidyl Metharylate (NGMA) polymer has been evaluated as a non-viral gene transfer platform by transferring plasmid DNA encoded with Green Fluorescent Protein (GFP). The NGMA–plasmid complexes were coated onto tissue culture polystyrene surfaces and L-929 cells were cultured on these surfaces. Gene transfer was induced through temperature modulation in the presence of a transfection reagent, enabling intracellular uptake and functional expression of GFP. The delivery of plasmids into the cell monolayer was evaluated in different conditions. Results demonstrated rapid and efficient cellular internalization of plasmid DNA, with distinct GFP expression observed only on polymer-coated regions, confirming the spatial and temporal control of this solid-state gene delivery system. This novel approach merges non-viral gene delivery with substrate-associated cell culture, providing a versatile platform for generating genetically modified cell monolayers.