Elucidating the interplay between DNA-condensing and free polycations in gene transfection through a mechanistic study of linear and branched PEI

Zhuojun Dai, Torben Gjetting, Maria Ahlm Mattebjerg, Chi Wu, Thomas Lars Andresen

    Research output: Contribution to journalJournal articleResearchpeer-review

    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 32P 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.
    Original languageEnglish
    JournalBiomaterials
    Volume32
    Issue number33
    Pages (from-to)8626-8634
    ISSN0142-9612
    DOIs
    Publication statusPublished - 2011

    Keywords

    • Polycations
    • PEI
    • Gene transfection
    • Non-viral vectors
    • Polyplex
    • Polyethylenimine

    Cite this