Gentle and efficient engineering of primary human NK cells by photoporation with polydopamine nanosensitizers

Abstract

Over the past several years, adoptive T cell therapies have accounted for great success in treating diverse malignancies. More recently, however, NK cells are being investigated as a promising alternative. Due to the innate antiviral properties of NK cells, viral engineering has proven to be challenging, prompting the development of non-viral transfection technologies. In this work, we evaluated photoporation with polydopamine nanosensitizers as a notable upcoming transfection technology for the engineering of NK cells and compared its performance to Nucleofection. Our results demonstrated the successful transfection of NK cells with eGFP mRNA and gene editing with Cas9 ribonucleoproteins (RNPs) for knock-out of the KLRC1 gene, encoding for the inhibitory NK cell receptor NKG2A. Importantly, no alterations to the phenotype of the cells (e.g. expression of surface markers and release of cytokines) could be detected, nor was the proliferation or cytolytic capacity of the cells influenced by either of the treatments. Overall, our findings highlight the potential of polydopamine-sensitized photoporation as a gentle and efficient transfection technology for NK cell engineering.