Abstract
Polyethylene glycol (PEG) functionalization of non-viral vectors represents a powerful tool through the formation of an overall surface charge shielding ability, which is fundamental for efficient nucleic acid delivery systems. The degree of non-viral vector PEGylation and the molecular weight of utilized PEG is crucial since the excessive use of PEG units may lead to a considerable reduction of the DNA-binding capacity and, subsequently, in a reduction of in vitro transfection efficiency. Herein, we report a detailed study on a series of dynamic combinatorial frameworks (DCFs) containing PEGylated squalene, poly-(ethyleneglycol)-bis(3-aminopropyl) of different lengths, and branched low molecular weight polyethylenimine components, reversibly connected in hyperbranched structures, as efficient dynamic non-viral vectors. The obtained frameworks were capable of forming distinct supramolecular amphiphilic architectures, shown by transmission electron microscopy (TEM) and dynamic light scattering (DLS), with sizes and stability depending on the length of PEG units. The interaction of PEGylated DCFs with nucleic acids was investigated by agarose gel retardation assay and atomic force microscopy (AFM), while their transfection efficiency (using pCS2+MT-Luc DNA as a reporter gene) and cytotoxicity were evaluated in HeLa cells. In addition, the data on the influence of the poly-(ethyleneglycol)-bis(3-aminopropyl) length in composition of designed frameworks over transfection efficiency and tolerance in human cells were analyzed and compared.
Highlights
Effective DNA delivery systems are multivalent “nanomachines”, with active functionalities integrated and positioned at the nanoscale [1]
Transfection tests were performed in three series: (i) NV1/pDNA-NV10/pDNA, A library of modular dynamic vectors with tuneable composition was prepared using earlier containing H2 N-Polyethylene glycol (PEG)-NH2 of 1500 Da in their compositions, starting with the ratio 0.1 equiv. for described protocols, containing hydrophobic component SQ-PEG-NH2, poly-(ethylene-glycol)-bis (3NV1 to 1 equiv. for NV10; (ii) NV11/pDNA-NV20/pDNA and NV21/pDNA-NV30/pDNA, containing amino-propyl)-terminated of various molecular weight (Mn 1500, 2000 and 3000 Da) NH2-PEG-NH2
For the NV11/pDNA-NV20/pDNA and from transmission electron microscopy (TEM) and dynamic light scattering (DLS) data that the increase in molecular weight of H2N-PEG-NH2 led to the formation
Summary
Effective DNA delivery systems are multivalent “nanomachines”, with active functionalities integrated and positioned at the nanoscale [1]. DNA and targeted cells are highly variable systems and rational design is limited to a relatively small number of components. Our group has been working on developing and implementing particular strategies to create a diversity of modular vectors capable of dynamically self-adapting to their DNA targets, allowing for the rapid screening of most effective vectors, optimally matched to DNA 3D surrounding space [6,7,8,9,10]. The use of reversible interactions as dynamic interfaces between the target and dynamical constitutional frameworks (DCF) components strongly contributed to the self-adjustment of the system’s tridimensional geometry and functional properties. A number of Polymers 2019, 11, 1313; doi:10.3390/polym11081313 www.mdpi.com/journal/polymers
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