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Lipid nanoparticles (LNPs), as utilized, e.g., in the COVID\u201019 vaccines, contain PEG\u2010lipids. However, alternatives are required because of the \u201cPEG dilemma\u201d recognized by an increase in anti\u2010PEG antibodies in the human population. In this study, poly(2\u2010ethyl\u20102\u2010oxazoline) (PEtOx)\u2010based lipids with different degrees of polymerization are synthesized and subsequently used to formulate mRNA\u2010loaded LNPs. The effect of polymer chain length on the size, immunoreaction, and transfection efficiency is investigated in detail. In addition, in\u2010depth transfection studies are performed using super\u2010resolution microscopy (SRM) to investigate the uptake mechanism of PEtOx\u2010based LNPs in comparison to PEG\u2010LNPs. 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