For mosquitoes the number of genetic modification technologies that can and have been applied continues to grow, and while still not as deep as the Drosophila toolbox the mosquito toolbox is still impressive in the right hands. Bernardini et al. nicely demonstrate three available technologies that they deftly apply to engineering the Y chromosome of Anopheles gambiae. They started with a fortuitous piggyBac transposon insertion on the Y chromosome, which also contained a meganuclease recognition site, 1-Sce1. Using that line they then used 1-Sce1 to create a double-stranded break in the Y chromosome to facilitate homologous recombination with a donor plasmid they introduced that contained a site-specific recombination site (attP) for phage C31 (phiC31). Taking this new line of An. gambiae they utilized the attP site to insert, by site-specific recombination, a marker gene.
So Bernardini et al. have strung together three technologies – transposon based integration, meganuclease based homologous recombination and site-specific recombination – to engineer the Y chromosome of An. gambiae. This was largely a demonstration of advanced insect genetic engineering in a challenging insect and presumably their Y-chromosome attP platform will allow them to either ‘make’ interesting insects for new genetics-based control strategies or to investigate the Y chromosome.
This is an Open Access article that is worth a look.
Bernardini F, Galizi R, Menichelli M, Papathanos P-A, Dritsou V, Marois E, Crisanti A, Windbichler N (2014) Site-specific genetic engineering of the Anopheles gambiae Y chromosome. Proceedings of the National Academy of Sciences 111: 7600-7605; doi:10.1073/pnas.1404996111