Optical tracking of organically modified silica nanoparticles as DNA carriers: a nonviral, nanomedicine approach for gene delivery.
This article reports a multidisciplinary approach to produce fluorescently labeled organically modified silica nanoparticles as a nonviral vector for gene delivery and biophotonics methods to optically monitor intracellular trafficking and gene transfection. Highly monodispersed, stable aqueous suspensions of organically modified silica nanoparticles, encapsulating fluorescent dyes and surface functionalized by cationic-amino groups, are produced by micellar nanochemistry. Gel-electrophoresis studies reveal that the particles efficiently complex with DNA and protect it from enzymatic digestion of DNase 1. The electrostatic binding of DNA onto the surface of the nanoparticles, due to positively charged amino groups, is also shown by intercalating an appropriate dye into the DNA and observing the Forster (fluorescence) resonance energy transfer between the dye (energy donor) intercalated in DNA on the surface of nanoparticles and a second dye (energy acceptor) inside the nanoparticles. Imaging by fluorescence confocal microscopy shows that cells efficiently take up the nanoparticles in vitro in the cytoplasm, and the nanoparticles deliver DNA to the nucleus. The use of plasmid encoding enhanced GFP allowed us to demonstrate the process of gene transfection in cultured cells. Our work shows that the nanomedicine approach, with nanoparticles acting as a drug-delivery platform combining multiple optical and other types of probes, provides a promising direction for targeted therapy with enhanced efficacy as well as for real-time monitoring of drug action.
Roy
Indrajit
I
Institute for Lasers, Photonics, and Biophotonics, University at Buffalo, State University of New York, Buffalo, NY 14260-3000, USA.
Ohulchanskyy
Tymish Y
TY
Bharali
Dhruba J
DJ
Pudavar
Haridas E
HE
Mistretta
Ruth A
RA
Kaur
Navjot
N
Prasad
Paras N
PN
D000818:Animals; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007624:KB Cells; D036103:Nanotechnology; D010957:Plasmids; D012822:Silicon Dioxide; D014162:Transfection / Q000379:methods*
D049452:Green Fluorescent Proteins; D012822:Silicon Dioxide; D004247:DNA