Highly efficient molecular delivery into mammalian cells using carbon nanotube spearing

Cai, Dong; Mataraza, Jennifer; Qin, Zheng‐Hong; Huang, Zhongping; Huang, Jianyu; Chiles, Thomas C.; Carnahan, David; Kempa, K.; Z, Ren

doi:10.1038/nmeth761

Cited by 526 publications

(373 citation statements)

References 24 publications

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“…The HEK293 cells adhered better on the CNT-loaded electrode than on the PEI/COOH-SAM. The CNTs clustering on the surface as seen in figure 2 might stick to the cell membrane (McKnight et al 2003) and thus gene transfer efficiency would increase (Cai et al 2005). The electron transfer between the gold thin film and immobilized CNTs has been reported even in the presence of the insulative SAM layer (Diao & Liu 2005).…”

Section: Resultsmentioning

confidence: 99%

Site-specific gene transfer with high efficiency onto a carbon nanotube-loaded electrode

Inoue

Fujimoto

Ogino

et al. 2008

J. R. Soc. Interface.

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A transfection array, which is specifically developed for use in high-throughput analyses of genome functions by the over-expression or suppression of genes on a chip, is expected to become an important method for post-genome research. High efficiency of gene expression or suppression is indispensable for high-throughput analyses because the adherent cell number on a single spot decreases as the density of spots increases in the transfection array. We have studied an electro-stimulated pore formation on the cell membrane for gene delivery. Fine pores should be formed on the cell membrane to increase the efficiency of gene transfection without cell damage. Herein, we examined the electrode carrying chemically functionalized carbon nanotubes (CNTs) on the surface. The CNTs were loaded on a gold electrode with a self-assembled monolayer membrane by electrostatic interaction. Adsorbed plasmid DNA was transfected with higher efficiency into adherent cells on the CNT-loaded electrode than on an electrode without CNTs. This result may be due to the strong but fine field emission formed from the tips of the CNTs.

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Section: Resultsmentioning

confidence: 99%

Site-specific gene transfer with high efficiency onto a carbon nanotube-loaded electrode

Inoue

Fujimoto

Ogino

et al. 2008

J. R. Soc. Interface.

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“…In many cases, uptake efficiency is reported with only a qualitative assessment of toxicity (67)(68)(69). Viral-derived peptides such as the TAT protein transduction domain have only been described as having "highly inefficient" uptake by neurons and "demonstrated toxicity" (70).…”

Section: Primary Antibodiesmentioning

confidence: 99%

“…Nearly 80% transduction of enhanced green fluorescent protein DNA was observed for carbon nanotube "spearing" in primary cortical neurons, but retraction of neurites also resulted from this invasive technique, which involves direct and physical penetration of neuronal cell membranes (69). Although recovery was said to have occurred by 48 h post-treatment, no toxicity data were given (69).…”

Section: Primary Antibodiesmentioning

confidence: 99%

Functional Protein Delivery into Neurons Using Polymeric Nanoparticles

Hasadsri

Kreuter

Hashimoto³

et al. 2009

Journal of Biological Chemistry

118

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An efficient route for delivering specific proteins and peptides into neurons could greatly accelerate the development of therapies for various diseases, especially those involving intracellular defects such as Parkinson disease. Here we report the novel use of polybutylcyanoacrylate nanoparticles for delivery of intact, functional proteins into neurons and neuronal cell lines. Uptake of these particles is primarily dependent on endocytosis via the low density lipoprotein receptor. The nanoparticles are rapidly turned over and display minimal toxicity to cultured neurons. Delivery of three different functional cargo proteins is demonstrated. When primary neuronal cultures are treated with recombinant Escherichia coli ␤-galactosidase as nanoparticle cargo, persistent enzyme activity is measured beyond the period of nanoparticle degradation. Delivery of the small GTPase rhoG induces neurite outgrowth and differentiation in PC12 cells. Finally, a monoclonal antibody directed against synuclein is capable of interacting with endogenous ␣-synuclein in cultured neurons following delivery via nanoparticles. Polybutylcyanoacrylate nanoparticles are thus useful for intracellular protein delivery in vitro and have potential as carriers of therapeutic proteins for treatment of neuronal disorders in vivo.

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“…The shape of the tube was elongated, suggesting a behavior as a nanoneedle, capable of piercing the lipid bilayer of the membrane without provoking cell death. This mechanism, which can be defined as nanopenetration, was observed by Cai and coworkers, who developed an efficient molecular delivery technique called nanotube spearing [192]. In this case, CNTs filled with magnetic particles were cultured with MCF-7 cells.…”

Section: Cnt Cell Uptakementioning

confidence: 69%