Combining sperm plug genotyping and coat color chimerism predicts germline transmission.
Combining sperm plug genotyping and coat color chimerism predicts germline transmission.
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DOI：
10.1007/s11248-013-9731-6
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发表时间：
2013-12
影响因子：
3
通讯作者：
Lloyd, K. C. Kent
中科院分区：
文献类型：
--
作者：
Lee, Angus Yiu-Fai;Evans, Kristin;Willis, Brandon;Lloyd, K. C. Kent
用于基因敲除小鼠的C57BL/ 6n来源的胚胎干细胞的使用显著增加。然而，在这种遗传背景下，嵌合体的种系传播（GLT）的潜力被观察到是高度可变的。利用毛色作为体细胞嵌合的指标来推断嵌合对生殖细胞群体的贡献程度，即使是高度agouti的C57BL/ 6n衍生的嵌合也可能无法实现GLT。我们研究了基因靶向胚胎干细胞中表达的标记基因的定量PCR基因分型在多大程度上可以对从交配塞中存在的精子中提取的DNA进行检测，以确定胚胎干细胞对生殖细胞的贡献。我们发现，对来自交配塞的精子DNA进行客观评估，并结合对毛色嵌合的主观评估，可以用来准确地为可能实现GLT的嵌合体的育种选择提供信息。这些结果表明，与随机选择嵌合体相比，包括对交配插头的分析来设置嵌合体进行繁殖有助于降低成本，减少时间，并促进需要嵌合体生产，选择，繁殖和测试以产生基因靶向小鼠的项目的研究。
There has been a significant increase in the use of C57BL/6N-derived ES cells for the production of gene knockout mice. However, the potential for germline transmission (GLT) from chimeras on this genetic background has been observed to be highly variable. Using coat color as an indicator of somatic chimerism to infer the extent of chimeric contribution to the germ cell population, even highly agouti C57BL/6N-derived chimeras can fail to achieve GLT. We investigated the extent to which quantitative PCR genotyping for a marker gene expressed in gene targeted ES cells can be performed on DNA extracted from sperm present in copulatory plugs to determine the contribution of ES cells to the germ cells. We found that an objective assessment of sperm DNA from copulatory plugs combined with a subjective assessment of coat color chimerism can be used to accurately inform the selection of chimeras for breeding that are likely to achieve GLT. These results indicate that, compared to random selection of chimeras, including an analysis of copulatory plugs to set chimeras for breeding can help to reduce costs, minimize time, and facilitate research for projects requiring the production, selection, breeding, and testing of chimeras to generate gene-targeted mice.
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影响因子：
64.8
作者：
Skarnes WC;Rosen B;West AP;Koutsourakis M;Bushell W;Iyer V;Mujica AO;Thomas M;Harrow J;Cox T;Jackson D;Severin J;Biggs P;Fu J;Nefedov M;de Jong PJ;Stewart AF;Bradley A
通讯作者：
Bradley A
影响因子：
2.7
作者：
Auerbach, W;Dunmore, JH;Joyner, AL
通讯作者：
Joyner, AL
影响因子：
7
作者：
Hansen, Gwenn M.;Markesich, Diane C.;Abuin, Alejandro
通讯作者：
Abuin, Alejandro
影响因子：
46.9
作者：
Poueymirou, William T.;Auerbach, Wojtek;Valenzuela, David M.
通讯作者：
Valenzuela, David M.
影响因子：
2.5
作者：
Hughes, Elizabeth D.;Qu, Yun Yan;Saunders, Thomas L.
通讯作者：
Saunders, Thomas L.