Ex vivo magnetofection: a novel strategy for the study of gene function in mouse organogenesis

Terje Svingen, Dagmar Wilhelm, Alexander N Combes, Brett Hosking, Vincent R Harley, Andrew H Sinclair, Peter A Koopman

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16 Citations (Scopus)


Gene function during mouse development is often studied through the production and analysis of transgenic and knockout models. However, these techniques are time- and resource-consuming, and require specialized equipment and expertise. We have established a new protocol for functional studies that combines organ culture of explanted fetal tissues with microinjection and magnetically induced transfection ( magnetofection ) of gene expression constructs. As proof-of-principle, we magnetofected cDNA constructs into genital ridge tissue as a means of gain-of-function analysis, and shRNA constructs for loss-of-function analysis. Ectopic expression of Sry induced female-to-male sex-reversal, whereas knockdown of Sox9 expression caused male-to-female sex-reversal, consistent with the known functions of these genes. Furthermore, ectopic expression of Tmem184a, a gene of unknown function, in female genital ridges, resulted in failure of gonocytes to enter meiosis. This technique will likely be applicable to the study of gene function in a broader range of developing organs and tissues.
Original languageEnglish
Pages (from-to)956 - 964
Number of pages9
JournalDevelopmental Dynamics
Issue number4
Publication statusPublished - 2009
Externally publishedYes

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