Improved <em>piggyBac</em> Transformation with Capped Transposase mRNA in Pest Insects

• DOI: 10.20944/preprints202308.2202.v1
• Publication Date: 2023-09-04T00:30:56Z
• Authors: Irina Haecker; Tanja Rehling; Henrik Schlosser; Daniela Mayorga-C...; Mara Heilig; Ying Yan; Peter Armbruster; Marc F. Schetelig
• Keywords: Transposase; Transgenesis

Creating transgenic insects is a key technology in insect genetics and molecular biology. A widely used instrument transgenesis the piggyBac transposase, resulting essentially ran-dom genomic integrations. In contrast, site-specific recombinases allow targeted integration of transgene construct into specific target site. Both strategies, however, often face limitations due to low rates. We aimed enhance rates by utilizing capped mRNA as source transposase or recombinase instead helper plasmid. system-atic comparison Aedes mosquitoes, models for hard transform in-sects, showed that suppling increased average transfor-mation efficiency aegypti from less than 5% with plasmid about 50% mRNA. Similar high transformation activity was observed Ae. albopictus pBac No differences between were recombination experi-ments. Furthermore, codon-optimized version delivered didn’t improve agricultural pest D. suzukii. believe use has strong potential enhancing efficiencies other mosquitoes important pests such tephritids.