title: Elucidating the interplay between DNA-condensing and free polycations in gene transfection through a mechanistic study of linear and branched PEI.
abstract: In the present study we compare LPEI and BPEI characteristics related to DNA condensation and their role as free polycation chains in gene transfection. Using radioactive (32)P labeled DNA, we investigated the effect of free PEI chains on the cellular uptake of polyplexes. Our investigations show different properties of BPEI and LPEI polyplexes in condensation and de-condensation processes as well as in cellular uptake, which was tightly correlated with transfection efficiency. In agreement with earlier reports we find all DNA to be condensed at N/P = 3. Further added PEI chains remain free in solution. We found that both the cellular uptake and gene transfection of BPEI polyplexes is much more efficient than LPEI polyplexes at a low N/P ratio of 3 (i.e., without free PEI chains). When N/P is high (10, with 7 portions of free PEI), the LPEI and BPEI polyplexes have similar transfection efficiency even though the cellular uptake of the LPEI polyplexes is significantly lower. In addition, we found that addition of free short or long PEI chains (2.5 and 25 kDa) leads to a comparable gene transfection efficiency.
authors.affiliation: Technical University of Denmark, DTU Nanotech, Department of Micro- and Nanotechnology, Building 423, 2800 Lyngby, Denmark.
authors.forename: Zhuojun
authors.initials: Z
authors.lastname: Dai
authors.forename: Torben
authors.initials: T
authors.lastname: Gjetting
authors.forename: Maria A
authors.initials: MA
authors.lastname: Mattebjerg
authors.forename: Chi
authors.initials: C
authors.lastname: Wu
authors.forename: Thomas L
authors.initials: TL
authors.lastname: Andresen
mesh_terms: D004247:DNA / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D011073:Polyamines; D000071228:Polyelectrolytes; D011094:Polyethyleneimine; D014162:Transfection
chemical_list: D004247:DNA; D011073:Polyamines; D000071228:Polyelectrolytes; D011094:Polyethyleneimine