High efficiency germ-line transformation of mosquitoes

Nature Protocols
Neil F LoboFrank H Collins

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.

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Oct 5, 2010·Cold Spring Harbor Protocols·Anthony ClemonsMolly Duman-Scheel
Oct 5, 2010·Cold Spring Harbor Protocols·Anthony ClemonsMolly Duman-Scheel
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Dec 4, 2020·PLoS Neglected Tropical Diseases·David Navarro-PayáLuke Alphey

Related Concepts

Genetic Techniques
Gametes
Microinjections
Culicidae
Transformation, Genetic
Recombinant Transgenes

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