A survey to establish performance standards for the production of transgenic mice
A survey to establish performance standards for the production of transgenic mice
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DOI：
10.1007/s11248-009-9335-3
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发表时间：
2010-08-01
影响因子：
3
通讯作者：
Montoliu, Lluis
中科院分区：
文献类型：
--
作者：
Fielder, Thomas J.;Barrios, Laura;Montoliu, Lluis
通过显微注射DNA到受精卵母细胞的原核中来产生转基因小鼠是在20世纪80年代早期描述的。很快就确定了影响该技术效率的许多参数，包括DNA构建体的类型，注射的DNA浓度，以及最重要的是，用于卵母细胞供体的小鼠品系。从那时起，世界各地的数百个实验室和转基因核心设施已经成功地使用这种技术，基本上如最初描述的那样，创造了数千种新的转基因小鼠品系。然而，就产生转基因小鼠所需的受精卵母细胞的数量而言，整个过程仍然相对低效，并且每天和构建体之间的产量变化可能很大。因此，核心设施往往很难向客户解释为什么没有从给定的构建体中产生足够数量的转基因创始人。我们认为，对平均产量和预期产量变化进行严格评估将使油田（和个别设施）受益。为此，我们从国际转基因技术学会（ISTT）网站（www.transtechsociety.org）发起了一项调查，以获得尽可能多的设施的原始显微注射数据。我们打算使用这些数据来建立该领域的性能标准。现有的设施将能够参考这些标准来处理不满意的客户，而新的设施将能够以可实现的目标为目标。我们甚至可以发现一个最佳的因素组合，使每一个设施，以实现更高的产量。
The generation of transgenic mice by microinjection of DNA into the pronuclei of fertilized oocytes was described in the early 1980s. A number of parameters affecting the efficiency of the technique were soon identified, including the type of DNA construct, the concentration of DNA being injected, and, most importantly, the strain of mice used for oocyte donors. Since then, hundreds of laboratories and transgenic core facilities across the world have successfully used this technique, essentially as originally described, to create thousands of new transgenic mouse lines. However, the overall procedure continues to be relatively inefficient, in terms of the number of fertilized oocytes required to produce a transgenic mouse, and variations in yields from day to day and construct to construct can be large. Consequently, core facilities often struggle to explain to their customers why a sufficient number of transgenic founders were not produced from a given construct. We believe the field (and individual facilities) would benefit from a rigorous assessment of average yields and expected variations in yields. To this end, we have initiated a survey from the International Society for Transgenic Technologies (ISTT) web site (www.transtechsociety.org), to obtain raw microinjection data from as many facilities as possible. We intend to use this data to establish performance standards for the field. Existing facilities will be able to refer to these standards in dealing with dissatisfied clients, and new facilities will be able to aim for an achievable goal. We may even be able to discover an optimum combination of factors that will allow every facility to achieve higher yields.