Abstract : Current transgenic methodology developed for mosquitoes has not been applied widely to the major malaria vector Anopheles gambiae, which has proved more difficult to genetically manipulate than other mosquito species and dipteran insects. In this protocol, we describe PhiC31-mediated site-specific integration of transgenes into the genome of A. gambiae. The PhiC31 system has many advantages over 'classical' transposon-mediated germline transformation systems, because it allows integration of large transgenes at specific, characterized genomic locations. Starting from a general protocol, we have optimized steps from embryo collection to co-injection of transgene-containing plasmid and in vitro-produced PhiC31 integrase mRNA. We also provide tips for screening transgenic larvae. The outlined procedure provides robust transformation in A. gambiae, resulting in homozygous transgenic lines in approximately 2-3 months.
https://hal-pasteur.archives-ouvertes.fr/pasteur-02008325 Contributor : Anne Lassailly-BondazConnect in order to contact the contributor Submitted on : Tuesday, February 5, 2019 - 4:11:51 PM Last modification on : Thursday, April 7, 2022 - 10:10:36 AM Long-term archiving on: : Monday, May 6, 2019 - 4:42:22 PM