Date of Award

8-2014

Degree Type

Masters Thesis

Degree Name

Master of Science (MS)

Department

Chemistry and Biochemistry

Committee Chair

Faqing Huang

Committee Chair Department

Chemistry and Biochemistry

Committee Member 2

Douglas Masterson

Committee Member 2 Department

Chemistry and Biochemistry

Committee Member 3

Yanlin Guo

Committee Member 3 Department

Biological Sciences

Abstract

Through layer-by-layer method, the authors have constructed three Au nanoplexes: AuPEI/RNA/PEI, AuPEI/RNA/PEI-mPEG, and AuPEI/NA/PEI-PEG-FA. All the nanoplexes are characterized by UV-vis spectrometry, DLS, and zeta potential. The surface density of the first layer PEI and second layer RNA were also determined. Moreover, the Au nanoplexes can protect siRNA from RNase degradation and are stable in cell culture medium.

siGLuc-ppp, ssRNA80, and siF17 were delivered by the assembled Au nanoplexes, and the results were analyzed by GLuc assay, TB assay, and Luciferase assay. In terms of RNA delivery, the Au nanoplexes AuPEI/RNA/PEI exhibited excellent gene silencing efficiency (or causing severe cell death) in the absence of the serum during the transfection. In terms of targeted delivery, PEGylation greatly reduces the nonspecific delivery of siRNA caused by AuPEI/siRNA/PEI. However, no apparent difference is found between AuPEI/RNA/PEI-mPEG and AuPEI/RNA/PEI-PEG-FA.

Fast fabrication, versatility, and ability for siRNA protection are the major characters of our new developed Au nanoplex system. The high gene silencing efficiency in the absence of serum makes it an excellent delivery agent for negative-charged macromolecules in vitro. However, the application of the system in targeted delivery needs more exploration.

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