High efficiency germ-line transformation of mosquitoes

Authors: 
Lobo NF, Clayton JR, Fraser MJ, Kafatos FC, Collins FH
Journal: 
Nat Protoc
Volume: 
1
Year: 
2006
Start Page: 
1 312
End Page: 
1 317
Issue: 
3
Database Issue: 
Abstract: 
The ability to manipulate the mosquito genome through germ-line transformation provides us with a powerful tool for investigating gene structure and function. It is also a valuable method for the development of novel approaches to combating the spread of mosquito-vectored diseases. To date, germ-line transformation has been demonstrated in several mosquito species. Transgenes are introduced into pre-blastocyst mosquito embryos using microinjection techniques that take a few hours, and progeny are screened for the presence of a marker gene. The microinjection protocol presented here can be applied to most mosquitoes and contains several improvements over other published methods that increase the survival of injected embryos and, therefore, the number of transformants. Transgenic lines can be established in approximately 1 month using this technique.