Pronuclear Injection

Since it was established in the early 1980s (1) pronuclear injection of fertilized mouse oocytes is the major technique used to generate genetically modified mice with a single integration site. Based on our previously published techniques (2-3), we have successfully generated many mouse models of diseases (4-6) and developed next generation inducible systems (7). The injection of a transgene into fertilized mouse oocytes is a straightforward approach that allows the generation of mutant founders in as little as 6 weeks. We offer microinjection of your transgene of interest and subsequent identification of founder animals by PCR.

Please contact us to discuss your strategy and timeline.

1) Gordon, J.W., Scangos, G.A., Plotkin, D.J., Barbosa, J.A., and Ruddle, F.H. (1980). Genetic transformation of mouse embryos by microinjection of purified DNA. Proceedings of the National Academy of Sciences of the United States of America 77, 7380-7384.

2) Ittner, L.M., and Gotz, J. (2007). Pronuclear injection for the production of transgenic mice. Nature protocols 2, 1206-1215.

3) Delerue, F. and Ittner, L.M. (2017). Generation of Genetically Modified Mice through the Microinjection of Oocytes. J Vis Exp. Jun 15;(124).

4) Ittner, L.M., Ke, Y.D., Delerue, F., Bi, M., Gladbach, A., van Eersel, J., Wolfing, H., Chieng, B.C., Christie, M.J., Napier, I.A., et al. (2010). Dendritic function of tau mediates amyloid-beta toxicity in Alzheimer's disease mouse models. Cell 142, 387-397.

5) Ittner A;Chua SW;Bertz J;Volkerling A;Van Der Hoven J;Gladbach A;Przybyla M;Bi M;Van Hummel A;Stevens CH;Ippati S;Suh LS;Macmillan A;Sutherland G;Kril JJ;Silva AP G;Mackay J;Poljak A;Delerue F;Ke YD;Ittner LM. (2016) 'Site-specific phosphorylation of tau inhibits amyloid-β toxicity in Alzheimer's mice', Science, vol. 354, pp. 904 - 908

6) Ke YD, van Hummel A, Stevens CH, Gladbach A, Ippati S, Bi M, Lee WS, Krüger S, van der Hoven J, Volkerling A, Bongers A, Halliday G, Haass NK, Kiernan M, Delerue F, Ittner LM. (2015) Short-term suppression of A315T mutant human TDP-43 expression improves functional deficits in a novel inducible transgenic mouse model of FTLD-TDP and ALS. Acta Neuropathol. Nov;130(5):661-78

7) Delerue F, White M, Ittner LM (2014). Inducible, tightly regulated and non-leaky neuronal gene expression in mice. Transgenic Research 23, 225-233.
 

Video: techniques used at the TAU to microinject mouse oocytes