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 Koopman
DEVELOPMENTAL DYNAMICS | WILEY | Published : 2009
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, where..View full abstract
Awarded by National Institutes of Health
Awarded by EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH &HUMAN DEVELOPMENT
Grant sponsor: Australian Research Council; Grant sponsor: National Health and Medical Research Council of Australia; Grant sponsor: National Institutes of Health; Grant number: HD049431.