Multilayer mediated forward and patterned siRNA transfection using linear-PEI at extended N/P ratios

Sumit Mehrotra, Ilsoon Lee, Christina Chan

    Research output: Research - peer-reviewArticle

    • 39 Citations

    Abstract

    Gene delivery from a substrate depends, in part, on the vector-nucleic acid complex that is bound to the surface and the cell adhesive properties of the surface. Here, we present a method to deliver patterns of small interfering RNA (siRNA) that capitalize on a forward transfection method (transfection by introducing siRNA transfection reagent complexes onto plated cells); herein denoted as multilayer mediated forward transfection (MFT). This method separates the substrate-mediated delivery from the cell adhesive properties of the surface. pH responsive layer-by-layer (LbL) assembled multilayers were used as the delivery platform and microcontact printing technique (μCP) was used to pattern nanoparticles of transfection reagent-siRNA complexes onto degradable multilayers. Efficient MFT depend on optimal formulation of the nanoparticles. 25 kDa linear polyethylenimine (LPEI) was optimized as the siRNA transfection reagent for normal forward transfection (NFT) of the nanoparticles. A broad range of LPEI-siRNA nitrogen/phosphate (N/P) ratios (ranging from 5 to 90) was evaluated for the relative amounts of siRNA incorporated into the nanoparticles, nanoparticle size and NFT efficiencies. All the siRNA was incorporated into the nanoparticles at N/P ratio near 90. Increasing the amount of siRNA incorporated into the nanoparticles, with increasing N/P ratio correlated with a linear blue shift in the ultraviolet/visible (UV/vis) absorbance spectrum of the LPEI-siRNA nanoparticles. NFT efficiency greater than 80% was achieved with minimal cytotoxicity at N/P ratio of 30 and siRNA concentration of 200 nM. Similarly, MFT efficiency ≥60% was achieved for LPEI-siRNA nanoparticles at N/P ratios greater than 30.

    LanguageEnglish (US)
    Pages1474-1488
    Number of pages15
    JournalActa Biomaterialia
    Volume5
    Issue number5
    DOIs
    StatePublished - Jun 2009

    Profile

    Polyetherimides
    Small Interfering RNA
    Multilayers
    RNA
    Transfection
    Nanoparticles
    Polyethyleneimine
    Adhesives
    Substrates
    Surface Properties
    Cytotoxicity
    Nucleic Acids
    Printing
    Nitrogen
    Genes
    Phosphates
    Nucleic acids

    Keywords

    • Layer-by-layer (LbL) assembly
    • LPEI-siRNA nanoparticles
    • N/P ratio
    • Patterned transfection
    • siRNA transfection

    ASJC Scopus subject areas

    • Biomaterials
    • Biomedical Engineering
    • Biotechnology
    • Biochemistry
    • Molecular Biology

    Cite this

    Multilayer mediated forward and patterned siRNA transfection using linear-PEI at extended N/P ratios. / Mehrotra, Sumit; Lee, Ilsoon; Chan, Christina.

    In: Acta Biomaterialia, Vol. 5, No. 5, 06.2009, p. 1474-1488.

    Research output: Research - peer-reviewArticle

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