Fig. 6: Spatial-selective delivery of mRNA by 200-NBs in adherent cells.

a Schematic representation of spatial-selective NB-mediated delivery of mRNA. b Confocal image 24 h after spatial-selective eGFP-mRNA transfection in HeLa cells according to a “J”-like pattern using 200-NBs at optimized conditions (1.3 × 10⁸ NBs/mL, 5 min incubation, 0.1 μg/μL eGFP mRNA). The bar represents 1000 µm. n = 3 images were recorded from three samples. c Cell-selective reprogramming with NBs by delivering CRE-mRNA in HeLa cells. (i) HeLa cells were previously transduced with a lentiviral construct containing the eFS-LoxP-DsRed Express II-rev(eGFP)-PxoL cassette. Untransfected cells will only express the red fluorescent protein DsRed Express II whereas in transfected cells with CRE mRNA the DsRed Express II-stop cassette will be inverted between the LoxP sites, resulting in eGFP expression. (ii) Confocal image 72 h after transfection of HeLa cells with CRE-mRNA using 200-NBs. Images were acquired in three spectral channels: Hoechst (nuclear stain, blue), DsRed Express II (red), and eGFP (green). The merged image obtained from the 3 channels it is shown in the last column. First row: control experiment where cells were incubated with mRNA and NBs but without laser irradiation. Second row: CRE mRNA transfection according to the “J”-like pattern. The bar represents 1000 µm. or each condition n = 3 images were recorded from three samples.