CRISPR/Cas9 Initiated Transgenic Silkworms as a Natural Spinner of Spider Silk
Class
Article
College
College of Science
Faculty Mentor
Randy Lewis Lewis
Presentation Type
Poster Presentation
Abstract
With a similar fiber spinning apparatus to spiders, silkworms are a potential host to spin spider silk-like fibers. In recent studies, it is still a challenge to incorporate and express native-size spider silk genes at a precise location in the genome of heterologous hosts. A fixed-point insertion and expression strategy has been designed in this study to overcome this limitation. Through CRISPR/Cas9 initiated non-homologous end joining, native-size spider silk genes (major ampullate spidroin 1 (MaSp1) and minor ampullate spidroin 1 (MiSp1) of Nephila clavipes) have been successfully integrated at defined locations in the fibroin heavy or light chain genes of silkworms. This is also the first report of using precise replacement through CRISPR/Cas9 and expressing native-size MaSp1 or MiSp1 using endogenous fibroin heavy/light chain promoters in silkworms. The transgenic spider/silkworm fibers exhibit improved mechanical properties approaching natural spider dragline silks. The strategy may also facilitate the integration and expression of large exogenous proteins at defined sites within a heterologous expression system.
Location
The North Atrium
Start Date
4-12-2018 10:30 AM
End Date
4-12-2018 11:45 AM
CRISPR/Cas9 Initiated Transgenic Silkworms as a Natural Spinner of Spider Silk
The North Atrium
With a similar fiber spinning apparatus to spiders, silkworms are a potential host to spin spider silk-like fibers. In recent studies, it is still a challenge to incorporate and express native-size spider silk genes at a precise location in the genome of heterologous hosts. A fixed-point insertion and expression strategy has been designed in this study to overcome this limitation. Through CRISPR/Cas9 initiated non-homologous end joining, native-size spider silk genes (major ampullate spidroin 1 (MaSp1) and minor ampullate spidroin 1 (MiSp1) of Nephila clavipes) have been successfully integrated at defined locations in the fibroin heavy or light chain genes of silkworms. This is also the first report of using precise replacement through CRISPR/Cas9 and expressing native-size MaSp1 or MiSp1 using endogenous fibroin heavy/light chain promoters in silkworms. The transgenic spider/silkworm fibers exhibit improved mechanical properties approaching natural spider dragline silks. The strategy may also facilitate the integration and expression of large exogenous proteins at defined sites within a heterologous expression system.