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bioRxiv · 10.1101/642413

Oligonucleotide Length Determines Intracellular Stability of DNA-Wrapped Carbon Nanotubes

Gravely, M.·Safaee, M. M.·Roxbury, D.
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Abstract

Non-covalent hybrids of single-stranded DNA and single-walled carbon nanotubes (SWCNTs) have demonstrated applications in biomedical imaging and sensing due to their enhanced biocompatibility and photostable, environmentally-responsive near-infrared (NIR) fluorescence. The fundamental properties of such DNA-SWCNTs have been studied to determine the correlative relationships between oligonucleotide sequence and length, SWCNT species, and the physical attributes of the resultant hybrids. However, intracellular environments introduce harsh conditions that can change the physical identities of the hybrid nanomaterials, thus altering their intrinsic optical properties. Here, through visible and NIR fluorescence imaging in addition to confocal Raman microscopy, we show that the oligonucleotide length determines the relative uptake, intracellular optical stability, and expulsion of DNA-SWCNTs in mammalian cells. While the absolute NIR fluorescence intensity of DNA-SWCNTs in murine macrophages increases with increasing oligonucleotide length (from 12 to 60 nucleotides), we found that shorter oligonucleotide DNA-SWCNTs undergo a greater magnitude of spectral shift and are more rapidly internalized and expelled from the cell after 24 hours. Furthermore, by labeling the DNA with a fluorophore that dequenches upon removal from the SWCNT surface, we found that shorter oligonucleotide strands are displaced from the SWCNT within the cell, altering the physical identity and changing the fate of the internalized nanomaterial. These findings provide fundamental understanding of the interactions between SWCNTs and live cells which can be applied towards development of robustly engineered carbon nanotube sensors while mitigating associated nanotoxicity.\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=80 SRC=\"FIGDIR/small/642413v1_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (36K):\norg.highwire.dtl.DTLVardef@f3b7bborg.highwire.dtl.DTLVardef@a36d0borg.highwire.dtl.DTLVardef@14c082corg.highwire.dtl.DTLVardef@acac73_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOTOC GraphicC_FLOATNO C_FIG

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Gravely, M., Safaee, M. M., Roxbury, D.. 2019-05-20. Oligonucleotide Length Determines Intracellular Stability of DNA-Wrapped Carbon Nanotubes. https://doi.org/10.1101/642413

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