Transgenic mosquitoes and Icilier bee molecules: can self-docking strains of Anopheles gambiae help engineer a malaria transmission blockade?

摘要

Transgenic mosquitoes are typically created using transposable elements even though this technology does have limitations. Transposons can only introduce relatively small sections of DNA and the random chromosomal location of integrated transgenes cancause insertional mutagenesis and position effects or variability on transgene expression. To circumvent these problems, we have pioneered the development of site-directed transgenesis using the phiC31 viral integrase (Nimmo et al., 2006; Meredith et al., 2011, 2013; Pondeville et al., 2014). Site-specificity arises from a two-phase approach. In phase 1, the docking site (phage attachment site - attP) is integrated into the genome using a standard transposable element. Many such docking strains can becreated and those that have the desired characteristics are chosen for subsequent experiments. In phase 2, which requires the presence of phiC31 integrase, the attP site in the docking strain accepts transgenes from any plasmid carrying the corresponding bacterial attachment site [attB], Recombination changes the attachment sites such that they are no longer recognized by the integrase, therefore insertions are efficient, unidirectional and stable.