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bioRxiv · 10.1101/2024.02.21.581498

Rapid generation and screening of transgenic black soldier fly (Hermetia illucens)

Pfitzner, C.·Tepper, K.·Kumar, S.·Retief, C.·McNab, J.·Harrell, R. A.·Maselko, M.
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Abstract

BackgroundThe black soldier fly (BSF), Hermetia illucens is a widely used, and mass-produced insect that fulfils an important role in both the management of organic waste and as a component of animal feed formulations. They also have significant potential as a platform for converting organic waste into high-value proteins, and lipids for the production of biofuels. Applying synthetic biology to BSF provides even more potential for improvement through the generation of transgenic BSF to enhance animal feed, produce and fine tune high-value industrial biomolecules, and to expand their waste conversion capabilities. ResultsTo enable the rapid generation and screening of transgenic BSF, we utilised microinjections of piggyBac mRNA with donor plasmids. We have found preliminary screening of G0 BSF to identify mosaics for outcrossing can be completed less than 2 weeks after microinjection. Stable transgenic lines were reliably generated with effective transformation rates of 30-33%, and transmission of the transgene could be confirmed 3 days after outcrossing the G0 adults. We also present a protocol for identifying the location of integrated transgenes. ConclusionsThe methods presented here expedite the screening process for BSF transgenesis and further expand the toolkit for BSF synthetic biology.

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Pfitzner, C., Tepper, K., Kumar, S., Retief, C., McNab, J., Harrell, R. A., Maselko, M.. 2024-02-22. Rapid generation and screening of transgenic black soldier fly (Hermetia illucens). https://doi.org/10.1101/2024.02.21.581498

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