Abstract
We investigated the application of cryopreserved pronuclear‐stage zygotes for the production of transgenic rats. Most of the pronuclear‐stage zygotes cryopreserved by conventional two‐step freezing or vitrification appeared morphologically normal, but the proportion of frozen zygotes that developed into fetuses following transfer (59.7–60.2%) was higher than that of vitrified zygotes (5.5–22.1%). When the frozen‐thawed zygotes were used for DNA microinjection, 97.5% survived after DNA microinjection and 25.1% of the transferred zygotes developed into fetuses. These proportions were comparable to those of the fresh control zygotes (97.0% and 30.0%, respectively). The integration efficiency of the exogenous DNA into fetuses was similar between the frozen group (3.3% per injected zygote) and the control group (3.5%). These results indicate that pronuclear‐stage rat zygotes can be successfully cryopreserved by conventional two‐step freezing for production of transgenic rats.
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Takahashi, R., Hirabayashi, M. & Ueda, M. Production of transgenic rats using cryopreserved pronuclear‐stage zygotes. Transgenic Res 8, 397–400 (1999). https://doi.org/10.1023/A:1008910629235
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DOI: https://doi.org/10.1023/A:1008910629235