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Pré-Publication, Document De Travail Année : 2023

Transfection of functional proteins with DNA-protein nanogels-lipids complex

Résumé

Using functional proteins for therapeutic purposes due to their high selectivity and/or catalytic properties can enable the control of various cellular processes, however, the transport of active proteins inside living cells remains a major challenge. In contrast, intracellular delivery of nucleic acids has become a routine method for a number of applications in gene therapy, genome-editing or immunization. Here we report a functionalizable platform constituting of DNA-protein nanogel carriers crosslinked through streptavidin-biotin interactions and demonstrate its applicability for intracellular delivery of active proteins. We demonstrate that the nanogels can be loaded with proteins bearing either biotin or streptavidin tags, and can be transfected into living cells after complexation with cationic lipid vectors. In particular we use this approach for transfection of alkaline phosphatase enzyme which is shown to keep its catalytic activity after internalization by mouse melanoma B16 cells, as demonstrated by DDAO-phosphate assay. The resulting functionalized nanogels have dimensions of the order of 100 nm, contain around 100 enzyme molecules and are shown to be transfectable at low lipid concentrations (charge ratio R+/- = 0.75). Low lipid concentrations requested for transfection of the 3-dimensional DNA nanogels ensure low toxicity of our system, which in combination with high local enzyme concentration (~ 100 µM) underlines potential interest of this nanoplatform for biomedical applications.
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Dates et versions

hal-04284754 , version 1 (14-11-2023)
hal-04284754 , version 2 (19-02-2024)

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Marina Mariconti, Virginie Escriou, Mathieu Morel, Damien Baigl, Céline Hoffmann, et al.. Transfection of functional proteins with DNA-protein nanogels-lipids complex. 2023. ⟨hal-04284754v1⟩
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