A nuclear targeting approach enables efficient transgenesis in the milkweed bug Oncopeltus fasciatus

Adding 1x and 3x nuclear localization signals to piggyBac mRNA substantially increases transformation efficiency for transgenesis in the milkweed bug Oncopeltus fasciatus.

• Why it matters: Species-specific barriers and the need for careful optimization limit gene editing and transgenesis in non-traditional insects, restricting comparative genetic studies.
• What they did: The authors used CRISPR/Cas9 to create an eye- and body-color mutant strain, then built a nuclear-targeted piggyBac transgenesis workflow and integrated cassettes from 1.8 to 7.2 kb.
• The result: They achieved functional phiC31 attP strains and histone-labelled live-imaging strains, and showed the Q binary expression system works in Oncopeltus fasciatus, enabling more complex genetic manipulations.

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