Uptake mechanisms of non-viral gene delivery

Development of a recombinant fusion protein based on the dynein light chain LC8 for non-viral gene delivery

“…Interestingly, the smallest complex which also showed no significant variation during the analysis was pDNA:LD4 (1:100) with Lipofectamine™. Particle size and particle surface charge are two main factors that dictates internalization of the delivery complex by the cell and is directly correlated to transfection efficiency [32,33]. Thus, by monitoring both variables we are able to correlate and perhaps explain the results found during transfection.…”

Parameters and characteristics governing cellular internalization and trans-barrier trafficking of nanostructures

“…Cellular internalization and trans-barrier trafficking of NPs Endocytic event cascade is activated by the signaling of the NP on the cell surface, 6 which aligns surface proteins to prompt clathrin recruitment from the cytosol to begin clathrin-coating on the inner membrane of the cell. An adaptor protein, Epsin, is involved in the initial stages of membrane curvature and pit formation and accessory proteins such as dynamin (GTPase) affect vesicle formation from shallow to deep invagination by inducing deformation of the membrane.…”

“…7 With the aid of dynamin, a clathrin-coated vesicle with a size of 100-150 nm is formed due to polymerization of the coat complex and the NP-containing clathrin-coated vesicle then internally detaches from the donor membrane. 6 Once within the cell, clathrin and adaptor proteins uncoat to allow fusing of the vesicle within the cell to release the endocytosed NPs ( Figure 3). …”

Low Molecular Weight Chitosan (LMWC)-based Polyplexes for pDNA Delivery: From Bench to Bedside

“…The usage of phagocytosis and macropinocytosis seems very limited, as it only exists in specialized cells such as macrophages monocytes, dendritic cells or antigen presenting cells. Thus, CME and CvME are the best-characterized types of endocytosis in non-viral gene therapy field, although CvME pathway is less well understood [56]. It is described that LMWC based polyplexes could enter the cell via these two endocytic pathways, however, there is a lack of consensus in the scientific area [53,57].…”

“…Regardless of the mechanism of internalization, polyplexes must escape from the endosome before being degraded in the lysosome. Polyplexes that enter the cell via CME are confined within endosomes that will suffer a maturation process involving the compartment acidification resulting in late endosomes and finally, lysosomes [56]. The release of the pDNA cargo from these vesicles seems to be the bottleneck in the transfection process [57], since the acidic environment inside lysosome may lead to the degradation of the pDNA if it does not escape from the endosome on time.…”