A non-surgical method for producing gene-modified mice

描述(由申请人提供)：我们建议开发一种新的非手术技术，使我们能够将胚胎和囊胚重新引入假孕雌性小鼠。这项技术基于多年前发表的方法，涉及通过阴道将细胞引入子宫。这些早期的方法使用玻璃管引入囊胚；我们开发了一种新的塑料装置，可以与P-2移液器一起使用，将胚胎和囊胚引入雌性小鼠阴道内。这个过程需要几秒钟，疼痛和不适程度很小，所以雌性不需要麻醉。 我们已经生成了一些初步数据，并表明我们可以将胚胎移植到输卵管中，其效率与外科手术相当。我们还证明，我们可以将这项技术用于胚胎聚集体来产生嵌合体小鼠。因此，这项新技术应该能够取代用于通过DNA显微注射和ES细胞介导的基因敲除来生产转基因小鼠的外科技术。我们相信，这是对目前方法的巨大改进，如果它真的像我们初步数据显示的那样成功，将成为转基因研究人员的首选方法。

DESCRIPTION (provided by applicant): We propose to develop a novel non-surgical technique that will allow us to reintroduce embryos and blastocysts into pseudopregnant female mice. This technology is based on methods that were published a number of years ago and involves the introduction of cells into the uterus by way of the vagina. These earlier methods used glass pipets to introduce blastocysts; we have developed a novel plastic device, which can be used with a P-2 pipetman, to introduce embryos and blastocysts intravaginally into female mice. This procedure takes several seconds and the pain and discomfort level is minimal so the females do not have to be anesthetized. We have generated some preliminary data and have shown that we can introduce embryos with an efficiency that is comparable to surgical transfer into the oviduct. We have also shown that we can use this technique with embryo aggregates to generate chimeric mice. Thus, this novel technology should be able to replace surgical techniques used for the production of genetically altered mice via DNA microinjection and ES-cell mediated gene knock-outs. We believe that this is a dramatic improvement over current methods, and would become adopted as the method of choice for transgenic researchers if it is truly as successful as our preliminary data suggests.