During the last decades, gene therapy has demonstrated its huge potential, attracting agrowing interest from the medicinal scientific community. However, it encounterslimitations due to the lack of suitable carriers to vectorize nucleic acids inside targetedcells. Nanogels are highly hydrated nano-size crosslinked polymeric networks that havebeen used in many biomedical applications, from drug delivery to tissue engineering anddiagnostics. Due to their easy production, tunability, and swelling properties they havecalled the attention as promising vectors for gene delivery. Precipitation polymerizationis a polymerization technique based on the difference of solubility between themonomers and the resulting growing polymers, largely reported in the preparation ofnanomaterials with advanced structures. Core-shell structures can be reached playingwith the monomers’ solubility and with the polymerization pathway. Dendriticpolyglycerol (dPG) is used as crosslinker, with N-isopropylacrylamide (NIPAM) and N-isopropylmethacrylamide (NIPMAM) as thermo-responsive monomers. Insights in thepolymerization mechanism are first reported. Aiming to introduce positive charges inthe core of such nanogels, two different set of nanogels are prepared from 2-dimethylaminothyl methacrylate (DMAEMA) and glycidyl methacrylate (GMA)copolymerization, with subsequent functionalization of GMA with ethylenediamine (ED)via the epoxy group. Then, the impact of DMAEMA and GMA copolymerization on thefinal structure using both batch and semi-batch pathways is investigated. Finally, thesecarriers are shown to be non-cytotoxic and can be loaded with plasmid and proteins.