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Tn5 Transposase-Mediated Mouse Transgenesis¹

Departament of Anatomy and Reproductive Biology,⁶ University of Hawaii School of Medicine, Honolulu, Hawaii 96822 Friedrich Miescher-Institute,⁷ 4058 Basel, Switzerland

ABSTRACT

We have developed a novel method for mouse transgenesis. The procedure relies on a hyperactive Tn5 transposase to insert a transgene into mouse chromosomes during intracytoplasmic sperm injection. This procedure integrates foreign DNA into the mouse genome with dramatically increased effectiveness as compared to conventional methods such as pronuclear microinjection and traditional sperm injection-mediated transgenesis. Our data indicate that with this method, transgenic mice, both hybrids and inbreds, can be produced more consistently and with lower numbers of manipulated oocytes required for traditional microinjection methods. The transposase-mediated transgenesis technique is also effective with round spermatids, offering the potential for rescuing the fertility of azoospermic animals using sperm precursor cells.

oocyte development, sperm, spermatid

¹ Supported by a Victoria S. & Bradley L. Geist Foundation grant to S.M. P.P., J.F.S., and B.H. were supported by the Novartis Research Foundation.

² Correspondence. FAX: 808 956 7316; moisyadi{at}hawaii.edu

³ These authors contributed equally to this work.

⁴ Current address: Department of Obstetrics and Gynecology, Fukushima Medical University, 1 Hikarigaoka, Fukushima, Japan 960-1295.

⁵ Current address: University of Zurich, Institute of Laboratory Animal Science, Sternwartstrasse 6, CH-8091 Zurich, Switzerland.

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