Date of Award:
Doctor of Philosophy (PhD)
Randolph V Lewis
Randolph V Lewis
Jon Y Takemoto
Ronald C Sims
James P. Pitts
Using silkworms as the potential host to spin spider silk-like fibers is an area of intense research world-wide. The conventional methods used to create transgenic silkworms hosting spider silk-like gene limits the incorporation of spider silk-like protein and do not improve the mechanical performance of the composite silkworm/spider silk fibers. In this dissertation, synthetic spider ampullate genes were incorporated into the precise site of the fibroin heavy chain or light chain using the latest genome editing technology CRISPR/cas9 guided non-homologous end joining as opposed to conventional random integration using transposon-based piggyBac system. These protocols, with extensive applicability to other silkworm researches, improved the content of spider silk-like protein in the transgenic silkworm/spider silk fibers, increases genetic stability in offspring, and improves the mechanical performance of the transgenic fibers compared to traditional methods. In addition, an enhanced green fluorescence protein (eGFP) was successfully incorporated into the fibroin light chain of silkworms using CRISPR/C as 9 initiated homologous recombination. The transgenic silkworm/spider fibers emitted strong green fluorescence under excitation. These results demonstrate that the we successfully developed a protocol to make silkworm as a host to spin spider silk-like fibers.
Zhang, Xiaoli, "Using Silkworms as a Host to Spin Spider Silk-Like Fibers" (2017). All Graduate Theses and Dissertations. 6368.
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