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Synthesis and Characterization of Zeolitic Imidazolate Framework-8 as a Carrier for Gene Therapy Applications

Montazeri, Farzad | 2021

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 54270 (07)
  4. University: Sharif University of Technology
  5. Department: Materials Science and Engineering
  6. Advisor(s): Simchi, Abdolreza; Behmanesh, Mehrdad
  7. Abstract:
  8. Advances in genetics and biology have led to the ability to produce therapeutic genes and modify defective genes in patients' genomes, resulting in a solution to many genetic diseases. Many genetic diseases can be overcome through gene therapy by transferring therapeutic genes into the cell nucleus. However, researchers have not yet been able to develop a safe, high-efficiency carrier for the transfer of genes to the target cell, and this has become a bottleneck for the clinical applications of gene therapy. In this study, the ZIF-8 organic-metallic framework was used as a new vector for the transfer of therapeutic genes to target cells. For this purpose, ZIF-8 was synthesized by a solvothermal method using 2 different solvents and after characterization, it was determined that synthesized ZIF-8, using water as a solvent, due to its higher specific surface area (about 120 m2), smaller particle size, and average size less than 200 nm, and better water biocompatibility, have better properties for use as a carrier compared to ZIF-8 synthesized with methanol. The ability of ZIF-8 to encapsulate plasmids was demonstrated using gel retardation assay. Then, using in situ synthesis, a ZIF-8 shell was created around the plasmids and plasmids have been protected in the extracellular environment. Upon entering the cell, ZIF-8 is degraded and the plasmid is released due to the acidic environment in the cytoplasm. Due to the presence of the GFP protein gene in the transported plasmid sequence, observation of green light under a fluorescent microscope ensured the correct transfer of the plasmids into the cell and the expression of the desired gene; That is, the plasmid encapsulated by ZIF-8, after entering the cell, is released from ZIF-8 and enters the nucleus of the cell and is then correctly expressed. The toxicity of the synthesized nanoparticles on 2 fibroblasts and N1 cell lines was investigated by alamarBlue method and it was observed that at concentrations less than 80 mM, cell viability is more than 90%. Compared to commercial gene carriers used to transfect cells in vitro, ZIF-8 has a much less defensible function that can be considered for commercialization
  9. Keywords:
  10. Plasmid ; Gene Delivery ; Metal-Organic Framework ; Zeolite Imidazolate Framework ; Gene Therapy

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