bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown
Опубликована: Ноя. 3, 2024
Abstract The development of natural or synthetic polycations able to interact with nucleic acids and condense them into nanoparticles known as polyplexes, faces several unresolved challenges at the cellular level. Key issues include intracellular trafficking endosomal escape release cytosol, which are considered major bottlenecks for efficient protein expression. Here, we aim gaining fundamental insights stability polyplexes in biological media their uptake trafficking, while correlating data expression reporter both molecular characteristics various poly(ethylenimines) (PEI) physicochemical PEI/peGFP-C3 polyplexes. For this, chosen four samples PEI, selected a model polycation, different weights (Mw = 0.8, 20, 25 60 kg/mol) structures (linear branched). We found that vitro vivo cell internalization transfection efficiency is dependent on variation polycation Mw structure, well intrinsic properties such charge ratio (R=[N + ]/[P − ]). A relation between percentage positive cells green fluorescent (GFP) amount internalized acid (cyanine 5-peGFP-C3) allowed revealing PEI promoting higher GFP HEK293T cells. In long term, outcome this work will be propose guidelines help design more effective, less cytotoxic non-viral gene carriers great potential new therapeutic applications.
Язык: Английский