Figure 1.

Schematic representation of the study design to achieve nucleic acid-directed co- production of target proteins in individual cells. Plasmid DNA was directly (co-)delivered to cells using an established carrier system (i.e. PEI) (upper panel) or used as a template to IVT-mRNA, followed by (co-)delivery of IVT-mRNA by LipoMM to cells (lower panel). The former requires nucleus entry and transcription to mRNA, whereas in the latter case IVT-mRNA is, upon cellular uptake and endosomal escape, instantly translated to protein in the cytoplasm (not illustrated here). In both cases, however, mRNA is the ultimate entity, which is processed by ribosomes as a blueprint for protein synthesis. The two distinct payload designs, namely monocistronic and bicistronic nucleic acid refer to two genes integrated within one continuous open reading frame, and different IVT-mRNAs co-formulated together in a single carrier type (e.g., lipoplex or polyplex) in a statistical fashion, respectively. Three different genes with distinct sizes were assessed as the first gene, while keeping EGFP as a fluorescent marker constantly as the second gene. The order of representation does not reflect the experimental sequence. (Illustration created by BioRender.com).