Characterization of Multilayered Nanoparticles Encapsulated in Yeast Cell Wall Particles for DNA Delivery

Nonviral gene delivery technologies have been developed using layer-by-layer self-assembly of nanomaterials held together by electrostatic interactions in order to provide nanoparticulate materials that protect and deliver DNA to cells. Here we report a new DNA delivery technology based on the in si...

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Bibliographic Details
Published in:Bioconjugate chemistry 2008-04, Vol.19 (4), p.840-848
Main Authors: Soto, Ernesto R, Ostroff, Gary R
Format: Article
Language:English
Subjects:
Animals
Cell Wall - chemistry
Cells
Deoxyribonucleic acid
DNA
DNA - metabolism
Flow Cytometry
Fluorescence
Gene Transfer Techniques - instrumentation
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Mice
Microscopy, Fluorescence
Nanoparticles
Nanoparticles - chemistry
Nanoparticles - toxicity
NIH 3T3 Cells
Polyethylene Glycols - chemistry
Polyethylene Glycols - metabolism
Porosity
Proteins
RNA, Transfer - chemistry
RNA, Transfer - metabolism
Saccharomyces cerevisiae - cytology
Spectrum analysis
Transfection
Transfer RNA
Yeast
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Summary:Nonviral gene delivery technologies have been developed using layer-by-layer self-assembly of nanomaterials held together by electrostatic interactions in order to provide nanoparticulate materials that protect and deliver DNA to cells. Here we report a new DNA delivery technology based on the in situ layer-by-layer synthesis of DNA nanoparticles caged within hollow yeast cell wall particles (YCWP). YCWP provide protection and facilitate oral and systemic receptor-targeted delivery of DNA payloads to phagocytic cells. The nanoparticles inside YCWP consist of a core of tRNA/polyethylenimine (PEI) followed by a DNA layer that is finally coated with a protective outer layer of PEI. Using fluorescein and rhodamine labeling of tRNA, PEI, and DNA, the layer-by-layer formation of the nanoparticles was visualized by fluorescent microscopy and quantitated by fluorescence spectroscopy and flow cytometry. Optimal conditions (tRNA:YCWP, PEI:YCWP ratios and DNA load levels) to synthesize YCWP encapsulated nanoparticles were determined from these results. The high in vitro transfection efficiency of this encapsulated DNA delivery technology was demonstrated by the transfection of NIH3T3-D1 cells with YCWP-tRNA/PEI/gWizGFP/PEI formulations containing low amounts of the plasmid gWizGFP per particle to maximally express green fluorescent protein (GFP).
ISSN:1043-1802
1520-4812
DOI:10.1021/bc700329p