Strain-dependent differences in the efficiency of transgenic mouse production
Strain-dependent differences in the efficiency of transgenic mouse production
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DOI：
10.1023/a:1022166921766
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发表时间：
2003-02-01
影响因子：
3
通讯作者：
Joyner, AL
中科院分区：
文献类型：
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作者：
Auerbach, AB;Norinsky, R;Joyner, AL
通过原核显微注射生产转基因小鼠是一个由许多连续步骤组成的复杂过程。许多不同的因素影响每个步骤的有效性，从而影响转基因小鼠生产的整体效率。卵子供体雌性对超排卵的反应、受精率、注射后卵子的存活率、受控胚胎植入和发育至足月的能力以及注射 DNA 的浓度和纯度都有助于转基因生产效率。我们使用四种不同的卵子供体小鼠品系评估并比较了转基因小鼠的生产效率：B6D2/F1 杂种、瑞士韦伯斯特 (SW) 远交、近交 FVB/N 和 C57BL/6。这些数据包括由高容量核心转基因小鼠设施进行的涉及类似 350 个 DNA 转基因构建体的实验。研究发现特定遗传背景对生产过程不同步骤的效率有显着影响。除了产蛋之外，FVB/N 小鼠在该过程的每个步骤中始终保持最高的转基因小鼠生产效率。 B6D2/F2 杂交卵也相当有效，但在 DNA 显微注射后比 FVB/N 卵裂解更频繁。另一方面，SW 卵在单细胞阶段比其他菌株的卵更容易阻塞。最后，使用 C57BL/6 蛋的主要限制因素是注射蛋产生的胎儿不像其他品系那样经常发育足月。根据我们的研究，可以针对每种卵供体品系修改转基因小鼠的生产程序，以克服任何缺陷，从而提高转基因小鼠生产的整体效率。
Transgenic mouse production via pronuclear microinjection is a complex process consisting of a number of sequential steps. Many different factors contribute to the effectiveness of each step and thus influence the overall efficiency of transgenic mouse production. The response of egg donor females to superovulation, the fertilization rate, egg survival after injection, ability of manipulated embryos to implant and develop to term, and concentration and purity of the injected DNA all contribute to transgenic production efficiency. We evaluated and compared the efficiency of transgenic mouse production using four different egg donor mouse strains: B6D2/F1 hybrids, Swiss Webster (SW) outbred, and inbred FVB/N and C57BL/6. The data included experiments involving similar to350 DNA transgene constructs performed by a high capacity core transgenic mouse facility. Significant influences of particular genetic backgrounds on the efficiency of different steps of the production process were found. Except for egg production, FVB/N mice consistently produced the highest efficiency of transgenic mouse production at each step of the process. B6D2/F2 hybrid eggs are also quite efficient, but lyze more frequently than FVB/N eggs after DNA microinjection. SW eggs on the other hand block at the 1-cell stage more often than eggs from the other strains. Finally, using C57BL/6 eggs the main limiting factor is that the fetuses derived from injected eggs do not develop to term as often as the other strains. Based on our studies, the procedure for transgenic mouse production can be modified for each egg donor strain in order to overcome any deficiencies, and thus to increase the overall efficiency of transgenic mouse production.