Enzymatically Ligated DNA–Surfactants: Unmasking Hydrophobically Modified DNA for Intracellular Gene Regulation |
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| Authors: | Alyssa K. Hartmann Dominic F. Cairns‐Gibson Joshua J. Santiana Mark Q. Tolentino Halle M. Barber Dr. Jessica L. Rouge |
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| Affiliation: | Department of Chemistry, University of Connecticut, Storrs, CT, USA |
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| Abstract: | Herein, we describe the characterization of a novel self‐assembling and intracellular disassembling nanomaterial for nucleic acid delivery and targeted gene knockdown. By using a recently developed nucleic acid nanocapsule (NAN) formed from surfactants and conjugated DNAzyme (DNz) ligands, it is shown that DNz–NAN can enable cellular uptake of the DNAzyme and result in 60 % knockdown of a target gene without the use of transfection agents. The DNAzyme also exhibits activity without chemical modification, which we attribute to the underlying nanocapsule design and release of hydrophobically modified nucleic acids as a result of enzymatically triggered disassembly of the NAN. Fluorescence‐based experiments indicate that the surfactant‐conjugated DNAzymes are better able to access a fluorescent mRNA target within a mock lipid bilayer system than the free DNAzyme, highlighting the advantage of the hydrophobic surfactant modification to the nucleic acid ligands. In vitro characterization of DNz–NAN's substrate‐cleavage kinetics, stability in biological serum, and persistence of knockdown against a proinflammatory transcription factor, GATA‐3, are presented. |
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| Keywords: | DNAzymes cleavage reactions gene knockdown mRNA surfactants |
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