Lipid nanoparticles (LNPs) have shown promise for delivering nucleic acids like DNA plasmids, but packaging large plasmids remains challenging. The findings and experimental work detailed in this thesis are geared towards providing a proof of concept and providing preliminary confirmation of the guiding principle.

We synthesized LNPs containing cationic lipids (D-Lin-MC3) to encapsulate the large pLVX-ZsGreen and psiCHECK (luciferase) plasmids (~10-14kb) using a microfluidic system. Characterization included size, charge, morphology, plasmid encapsulation efficiency and in vitro/in vivo studies. The LNP-pDNA particles were ~100nm in size with high encapsulation efficiency. The LNPs efficiently transfected 293T cells with low cytotoxicity, enabling strong expression of the ZsGreen and luciferase reporter genes. However, there were limitations to detect luciferase expression by bioluminescence imaging in mice after systemic LNP-pDNA administration. In conclusion, our results supported hypothesis. LNPs can package large plasmid DNA while maintaining favorable physicochemical properties and in vitro transfection, but in vivo delivery requires further optimization to achieve sufficient gene expression.

Date of publication

Spring 5-29-2024

Document Type




Persistent identifier



Masters in Biotechnology

Included in

Nanomedicine Commons