Production of transgenic rats using cryopreserved pronuclear‐stage zygotes
Production of transgenic rats using cryopreserved pronuclear‐stage zygotes
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使用冷冻保存的原核阶段受精卵生产转基因大鼠
DOI：
10.1023/a:1008910629235
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发表时间：
1999
影响因子：
3
通讯作者：
M. Ueda
中科院分区：
文献类型：
--
作者：
R. Takahashi;M. Hirabayashi;M. Ueda
我们研究了冷冻保存的原核期受精卵在转基因大鼠生产中的应用。大多数通过常规两步冷冻或玻璃化冷冻保存的原核期受精卵在形态上看起来正常，但移植后发育成胎儿的冷冻受精卵比例（59.7-60.2%）高于玻璃化受精卵（5.5-22.1%）。当冻融的受精卵用于DNA显微注射时，97.5%的受精卵在DNA显微注射后存活，25.1%的转移的受精卵发育成胎儿。这些比例与新鲜对照合子的比例相当（分别为97.0%和30.0%）。冷冻组（3.3%/注射受精卵）和对照组（3.5%）之间外源DNA整合到胎儿中的效率相似。这些结果表明，原核期大鼠受精卵可以通过常规的两步冷冻成功地冷冻保存，用于生产转基因大鼠。
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.