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Self-Assembly DNA nanogel with embedded functional proteins for Nanomedicine platform

  • Author(s): Lee, Jinhyung
  • Advisor(s): Chen, Yi
  • et al.
Abstract

A platform is established to embed functional proteins onto a DNA nanogel. These proteins are embedded through self-assembly mechanisms between the DNA and the proteins. The DNA nanogel is composed of single stranded DNAs, which forms a 3-dimensional nanogel structure by using a G-Quadruplex secondary structure as the crosslinker. The DNA nanogel has multiple overhanging 20 adenosine sequences, which can be used to modify the nanogels by simple Watson-Crick base pairing. In this thesis, we have demonstrated that the DNA nanogel has embedded the unmodified proteins while still preserving the structure and function of the proteins. As a proof of concept, we have confirmed the embedding with mobility shift assay of native agarose gel electrophoresis, imaged embedded proteins onto DNA nanogel with Atomic Force Microscopy(AFM), characterized the size and zeta potential with Dynamic Light Scattering(DLS). In addition, we have proved that embedded proteins preserved their function by showing the nanogel-IgG system can be conjugated to the surface proteins of E.Coli. To develop our system as a Nanomedicine platform, we have modified our nanogel to be used as targeted drug delivery vehicle for KB cells. Doxorubicin was loaded onto the DNA nanogel with embedded Immunoglobulin G(IgG)s. The nanogel-IgG system showed higher cytotoxicity to KB cells than controls due to the ability of the IgG to function as a targeting ligand. This result demonstrates the self-assembled system can function as a possible next generation drug delivery vehicle.

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