Gelatin nanofiber mats with Lipofectamine/plasmid DNA complexes for in vitro genome editing

Gelatin electrospun nanofiber mats are gaining interest for applications in biomaterials science, such as tissue engineering and drug/gene delivery systems. In this study, we report the use of electrospun gelatin nanofiber mats for plasmid DNA (pDNA) delivery. Gelatin nanofiber mats were insolubilized via cross-linking with glutaraldehyde. On the cross-linked mats, human embryonic kidney-derived HEK293 cells demonstrated high viability for 7 days of culture (>95%) and were able to proliferate during that time. The Lipofectamine/pDNA complexes were immobilized on the mats through immersion in a solution, and HEK293 cells cultured on these mats expressed GFP for 7 days. Furthermore, HEK293 cells did not express GFP via the pDNA complexes released from the mats because the ability to deliver pDNA into the cells was lost. Since the mats could be used to transfect multiple types of pDNA into the cells simultaneously, we have achieved targeted genome editing using the mats. These data highlight the potential of gelatin nanofiber mats with Lipofectamine/pDNA complexes for local gene therapy via pDNA delivery as well as genome editing.