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

  1. Terje Svingen1,†,
  2. Dagmar Wilhelm1,†,
  3. Alexander N. Combes1,
  4. Brett Hosking1,
  5. Vincent R. Harley2,
  6. Andrew H. Sinclair3,
  7. Peter Koopman1,*

Article first published online: 19 MAR 2009

DOI: 10.1002/dvdy.21919

Issue

Developmental Dynamics

Developmental Dynamics

Volume 238, Issue 4, pages 956–964, April 2009

Additional Information

How to Cite

Svingen, T., Wilhelm, D., Combes, A. N., Hosking, B., Harley, V. R., Sinclair, A. H. and Koopman, P. (2009), Ex vivo magnetofection: A novel strategy for the study of gene function in mouse organogenesis. Dev. Dyn., 238: 956–964. doi: 10.1002/dvdy.21919

Author Information

  1. 1

    Division of Molecular Genetics and Development, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Australia

  2. 2

    Human Molecular Genetics Laboratory, Prince Henry's Institute of Medical Research, Clayton, Australia

  3. 3

    Murdoch Children's Research Institute and Department of Paediatrics, The University of Melbourne, Royal Children's Hospital, Melbourne, Australia

  1. †

    Drs. Svingen and Wilhelm contributed equally to this work.

*Division of Molecular Genetics and Development, Institute for Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia

  1. †

    Drs. Svingen and Wilhelm contributed equally to this work.

Publication History

  1. Issue published online: 19 MAR 2009
  2. Article first published online: 19 MAR 2009
  3. Manuscript Accepted: 4 FEB 2009

Funded by

  • Australian Research Council
  • National Health and Medical Research Council of Australia
  • National Institutes of Health. Grant Number: HD049431

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Keywords:

  • gene delivery;
  • sex determination;
  • germ cells;
  • meiosis;
  • Tmem184a;
  • Sry;
  • Sox9

Abstract

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. Developmental Dynamics 238:956–964, 2009. © 2009 Wiley-Liss, Inc.

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