Gold nanorod--siRNA induces efficient in vivo gene silencing in the rat hippocampus.

AIM Gold nanorods (GNRs), cellular imaging nanoprobes, have been used for drug delivery therapy to immunologically privileged regions in the brain. We demonstrate that nanoplexes formed by electrostatic binding between negatively charged RNA and positively charged GNRs, silence the expression of the target housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) within the CA1 hippocampal region of the rat brain, without showing cytotoxicity. MATERIALS & METHODS Fluorescence imaging with siRNA(Cy3)GAPDH and dark-field imaging using plasmonic enhanced scattering from GNRs were used to monitor the distribution of the nanoplexes within different neuronal cell types present in the targeted hippocampal region. RESULTS & CONCLUSION Our results show robust nanoplex uptake and slow release of the fluorescent gene silencer with significant impact on the suppression of GAPDH gene expression (70% gene silencing, >10 days postinjection). The observed gene knockdown using nanoplexes in targeted regions of the brain opens a new era of drug treatment for neurological disorders.

[1]  Paras N. Prasad,et al.  Introduction to Biophotonics , 2003 .

[2]  Ying Liu,et al.  Surface chemistry and aspect ratio mediated cellular uptake of Au nanorods. , 2010, Biomaterials.

[3]  Adela C. Bonoiu,et al.  Gold nanorod delivery of an ssRNA immune activator inhibits pandemic H1N1 influenza viral replication , 2010, Proceedings of the National Academy of Sciences.

[4]  Yu-cheng Tseng,et al.  Biodegradable calcium phosphate nanoparticle with lipid coating for systemic siRNA delivery. , 2010, Journal of controlled release : official journal of the Controlled Release Society.

[5]  Mark E. Davis,et al.  Evidence of RNAi in humans from systemically administered siRNA via targeted nanoparticles , 2010, Nature.

[6]  M. El-Sayed,et al.  Nuclear targeting of gold nanoparticles in cancer cells induces DNA damage, causing cytokinesis arrest and apoptosis. , 2010, Journal of the American Chemical Society.

[7]  Leaf Huang,et al.  Nanoparticles evading the reticuloendothelial system: role of the supported bilayer. , 2009, Biochimica et biophysica acta.

[8]  Hong Ding,et al.  Biocompatible near-infrared quantum dots as ultrasensitive probes for long-term in vivo imaging applications. , 2009, Small.

[9]  A. Bhattacharjee,et al.  cAMP-dependent kinase does not modulate the Slack sodium-activated potassium channel , 2009, Neuropharmacology.

[10]  P. Sharp,et al.  Functional Delivery of siRNA in Mice Using Dendriworms , 2009, ACS nano.

[11]  E. Marbán,et al.  Targeted knock-down of neuronal nitric oxide synthase expression in basal forebrain with RNA interference , 2009, Journal of Neuroscience Methods.

[12]  Adela C. Bonoiu,et al.  Nanotechnology approach for drug addiction therapy: Gene silencing using delivery of gold nanorod-siRNA nanoplex in dopaminergic neurons , 2009, Proceedings of the National Academy of Sciences.

[13]  Mark E. Davis.  The first targeted delivery of siRNA in humans via a self-assembling, cyclodextrin polymer-based nanoparticle: from concept to clinic. , 2009, Molecular pharmaceutics.

[14]  D. Thakker,et al.  Intracerebroventricular amyloid-β antibodies reduce cerebral amyloid angiopathy and associated micro-hemorrhages in aged Tg2576 mice , 2009, Proceedings of the National Academy of Sciences.

[15]  P. Prasad,et al.  Preparation of Gold Nanoparticles and their Applications in Anisotropic Nanoparticle Synthesis and Bioimaging , 2009 .

[16]  Hong Ding,et al.  Metallic Nanostructures as Localized Plasmon Resonance Enhanced Scattering Probes for Multiplex Dark Field Targeted Imaging of Cancer Cells. , 2009, The journal of physical chemistry. C, Nanomaterials and interfaces.

[17]  Vincent M Rotello,et al.  Gold nanoparticles in delivery applications. , 2008, Advanced drug delivery reviews.

[18]  M. Stachowiak,et al.  Nonviral gene transfection nanoparticles: function and applications in the brain. , 2008, Nanomedicine : nanotechnology, biology, and medicine.

[19]  Rebecca Richards-Kortum,et al.  Aptamer-Targeted Gold Nanoparticles As Molecular-Specific Contrast Agents for Reflectance Imaging , 2008, Bioconjugate chemistry.

[20]  M. Swihart,et al.  Templated Synthesis of Gold Nanorods (NRs): The Effects of Cosurfactants and Electrolytes on the Shape and Optical Properties , 2008 .

[21]  Arezou A Ghazani,et al.  Assessing the effect of surface chemistry on gold nanorod uptake, toxicity, and gene expression in mammalian cells. , 2008, Small.

[22]  L. Sui,et al.  Down-regulation of β1-adrenoceptors gene expression by short interfering RNA impairs the memory retrieval in the basolateral amygdala of rats , 2007, Neuroscience Letters.

[23]  P. Jain,et al.  Gold nanoparticles: interesting optical properties and recent applications in cancer diagnostics and therapy. , 2007, Nanomedicine.

[24]  I. Roy,et al.  ORMOSIL nanoparticles as a non-viral gene delivery vector for modeling polyglutamine induced brain pathology , 2007, Journal of Neuroscience Methods.

[25]  Hong Ding,et al.  Gold Nanorods Coated with Multilayer Polyelectrolyte as Contrast Agents for Multimodal Imaging , 2007 .

[26]  Mark E. Davis,et al.  Effect of siRNA nuclease stability on the in vitro and in vivo kinetics of siRNA‐mediated gene silencing , 2007, Biotechnology and bioengineering.

[27]  Joseph Irudayaraj,et al.  Identity profiling of cell surface markers by multiplex gold nanorod probes. , 2007, Nano letters.

[28]  W. Pardridge.  shRNA and siRNA delivery to the brain. , 2007, Advanced drug delivery reviews.

[29]  Daniel Hoyer,et al.  RNA interference for studying the molecular basis of neuropsychiatric disorders. , 2007, Current opinion in drug discovery & development.

[30]  Shan Jiang,et al.  Quantum-dot based nanoparticles for targeted silencing of HER2/neu gene via RNA interference. , 2007, Biomaterials.

[31]  L. Chau,et al.  Sensing capability of the localized surface plasmon resonance of gold nanorods. , 2007, Biosensors & bioelectronics.

[32]  J. Krueger,et al.  TNFalpha siRNA reduces brain TNF and EEG delta wave activity in rats. , 2007, Brain Research.

[33]  R. Juliano,et al.  Tat-Conjugated PAMAM Dendrimers as Delivery Agents for Antisense and siRNA Oligonucleotides , 2005, Pharmaceutical Research.

[34]  John F. Cryan,et al.  Model organisms: The ascent of mouse: advances in modelling human depression and anxiety , 2005, Nature Reviews Drug Discovery.

[35]  E. Stachowiak,et al.  Organically modified silica nanoparticles: a nonviral vector for in vivo gene delivery and expression in the brain. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[36]  P. Whittaker,et al.  RNA interference: From gene silencing to gene-specific therapeutics , 2005, Pharmacology & Therapeutics.

[37]  C. Murphy,et al.  Gold nanoparticles are taken up by human cells but do not cause acute cytotoxicity. , 2005, Small.

[38]  M. Ferrari.  Cancer nanotechnology: opportunities and challenges , 2005, Nature Reviews Cancer.

[39]  George M Whitesides,et al.  Nanoscience, nanotechnology, and chemistry. , 2005, Small.

[40]  F. Natt,et al.  Neurochemical and behavioral consequences of widespread gene knockdown in the adult mouse brain by using nonviral RNA interference. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[41]  W. Siegert,et al.  Guideline to reference gene selection for quantitative real-time PCR. , 2004, Biochemical and biophysical research communications.

[42]  S A Bustin,et al.  Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. , 2002, Journal of molecular endocrinology.

[43]  C. R Bjarkam,et al.  New strategies for embedding, orientation and sectioning of small brain specimens enable direct correlation to MR-images, brain atlases, or use of unbiased stereology , 2001, Journal of Neuroscience Methods.

[44]  R. Spengler,et al.  Neuronal-associated tumor necrosis factor (TNFα): its role in noradrenergic functioning and modification of its expression following antidepressant drug administration , 1997, Journal of Neuroimmunology.

[45]  V. Debabov.  Biotechnology and Chemistry , 1991 .

[46]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[47]  G. Paxinos,et al.  The Rat Brain in Stereotaxic Coordinates , 1983 .