Immobilization and release strategies for DNA delivery using carbon nanofiber arrays and self-assembled monolayers.
We report a strategy for immobilizing dsDNA (double-stranded DNA) onto vertically aligned carbon nanofibers and subsequently releasing this dsDNA following penetration and residence of these high aspect ratio structures within cells. Gold-coated nanofiber arrays were modified with self-assembled monolayers (SAM) to which reporter dsDNA was covalently and end-specifically bound with or without a cleavable linker. The DNA-modified nanofiber arrays were then used to impale, and thereby transfect, Chinese hamster lung epithelial cells. This mechanical approach enables the transport of bound ligands directly into the cell nucleus and consequently bypasses extracellular and cytosolic degradation. Statistically significant differences were observed between the expression levels from immobilized and releasable DNA, and these are discussed in relation to the distinct accessibility and mode of action of glutathione, an intracellular reducing agent responsible for releasing the bound dsDNA. These results prove for the first time that an end-specifically and covalently SAM-bound DNA can be expressed in cells. They further demonstrate how the choice of immobilization and release methods can impact expression of nanoparticle delivered DNA.
Materials Science and Technology Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6030, USA. peckysdb@ornl.gov
Diana B
DB
Peckys
Anatoli V
AV
Melechko
Michael L
ML
Simpson
Timothy E
TE
McKnight
D000818:Animals; D002244:Carbon / Q000378:metabolism*; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D043942:Nanotubes / Q000737:chemistry*; D014162:Transfection / Q000379:methods*
D002244:Carbon; D004247:DNA