Impalefection

Impalefection is a method of gene delivery using nanomaterials, such as carbon nanofibers, nanotubes, nanowires. Carbon nanotubes, owing to their characteristic strength and unique electrical properties, have found utility as scanning probe tips, solar cells and even bikes for the Tour de France. More recently, the multifunctional material has been used to directly introduce genetic material into cells. Previously deemed ‘impalefection’ by McKnight et al., this technique of cell transfection uses DNA-appended nanofibers to directly introduce DNA by piercing the cell. Peckys et al. report an approach to immobilize and release nanoparticle-bound DNA through impalefection using vertically-aligned carbon nanofiber arrays. DNA was chemisorbed onto gold-coated nanofiber arrays or conjugated to the gold-coated nanofibers with the cleavable linker, 3([2-aminoethyl]dithio)propionic acid or the linker, 6-aminocaproic acid. Fluorescent staining of DNA first used to evaluate the binding and release of the DNA when the 3([2-aminoethyl]dithio)propionic acid linker was reduced by glutathione. Chinese hamster lung fibroblast cells were pressed onto the nanofiber arrays by centrifugation, enabling direct penetration of DNA. The immobilization, delivery and expression of an in vitro transfection of the cells were assessed using epifluorescent imaging of yellow fluorescent protein expression. Impalefection was determined to be a viable approach for DNA transfection and subsequent protein translation. Yellow fluorescent protein expression levels in the lung cells showed some transfection of DNA covalently attached to the nanofiber arrays, but greater enhancement appeared when the DNA was attached to the nanofiber arrays with glutathione reducible linkers for subsequent release. This method of direct DNA delivery into a cell’s nucleus may eliminate or reduce the need for intracellular uptake enhancement or degradation protection. Future studies may involve the testing of other DNA molecules to show a range of protein translation. Many transfection methods are available and impalefection should be compared to determine potential advantages. This particular method offers a unique means of transfection that appears to bypass extracellular and cytosolic degradation using molecular immobilization and release strategies in conjunction with nanofiber arrays.

Further information

1. Tracking gene expression after DNA delivery using spatially indexed nanofiber arrays McKnight, T.E., A.V. Melechko, D.K. Hensley, D.G.J. Mann, G.D. Griffin, and M.L. Simpson, Nano Letters, 2004. 4(7): p. 1213-1219.
2. Bellucci S: Carbon nanotubes: physics and applications. Phys. Stat. Sol.2, 34–47 (2005).
3. Peckys DB, Melechko AV, Simpson ML, McKnight TE: Immobilization and release strategies for DNA delivery using carbon nanofiber arrays and self-assembled monolayers. Nanotechnology20, 145304–145312 (2009).
4. McKnight TE, Melechko AV, Hensley DK, Mann DGJ, Griffin DG, Simpson ML: Tracking gene expression and DNA delivery using spatially indexed nanofiber arrays. Nano Lett.4, 1213–1219 (2004).
5. «An Attempt of Nanowire Aided Impalefection of Nerve Cells». Louise Bohlin, Project for FYS 241, Experimental Biophysics, May 2006, Lund University, Sweden
6. Статья Impalefection из Wikipedia, свободной энциклопедии. Доступно под лицензией Creative Commons Attribution-Share Alike.

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