Animal & Dairy Sciences

Auburn University

ANSC 362
Repro-Tech Update - 12

Mammalian Transgenesis by Intracytoplasmic Sperm Injection

Anthony C. F. Perry, Teruhiko Wakayama, Hidefumi Kishikawa,
Tsuyoshi Kasai, Masaru Okabe, Yutaka Toyoda, Ryuzo Yanagimachi
Department of Anatomy and Reproductive Biology
University of Hawaii, Honolulu, HI

Press Release of May 14, 1999
Also See: Science, Vol 284, No. 5417, Pp. 1180-1183

May 14, 1999 / The American Association for the Advancement of Science / Volume 284, Number 5417 / -- Coinjection of unfertilized mouse oocytes with sperm heads and exogenous DNA encoding either a green fluorescent protein (GFP) or [beta]-galactosidase reporter produced 64 to 94 percent transgene-expressing embryos, reflecting DNA-sperm head association before coinjection. Nonselective transfer to surrogate mothers of embryos in the GFP series generated about 20 percent offspring expressing the integrated transgene. These data indicate that exogenous DNA can reproducibly be delivered into an oocyte by microinjected spermatozoa and suggest an adaptable method of transgenesis.

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Figure. 2. Transgenic embryos produced by single-shot double transgenesis. Oocytes were microinjected with spermatozoa that had been preincubated with a mixture of pCX--LacZ and pCX-EGFP tg DNAs. The same embryos are shown (X400 [original]) after 3.5 days viewed by Hoffman modulation contrast microscopy unstained (A; top), for GFP expression under long-wavelength (480 nm) UV light (B; middle), and stained with X-gal for beta-galactosidase expression (C).
[Reproduced from: Perry et al. 1999. SCIENCE 284, P.1181, Fig. 2.]