Coutelle Charles, Waddington Simon N, Themis Michael
National Heart and Lung Institute, Molecular and Cellular Medicine Section, Imperial College London, London, UK.
Methods Mol Biol. 2012;891:329-40. doi: 10.1007/978-1-61779-873-3_15.
So far no systematic studies have been conducted to investigate developmental aberrations after prenatal gene transfer in mice. Here, we suggest procedures for such observations to be applied, tested and improved in further in utero gene therapy experiments. They are based on our own experience in husbandry for transgenic human diseases mouse models and breading, rearing, and observing mice after fetal gene transfer as well as on the systematic screens for monitoring of knock-out mutant mouse phenotypes established in international mutagenesis projects (EUMORPHIA and EUMODIC and subsequently the International Mouse Phenotyping Consortium). We also describe here the analysis procedures for detection of germ line mutations based on quantitative PCR (qPCR) by sperm-DNA analysis and breeding studies.
到目前为止，尚未进行系统研究来调查小鼠产前基因转移后的发育异常情况。在此，我们提出了一些程序，以便在进一步的子宫内基因治疗实验中应用、测试和改进此类观察。这些程序基于我们在转基因人类疾病小鼠模型的饲养、繁殖、饲养和观察胎儿基因转移后的小鼠方面的经验，以及基于国际诱变项目（EUMORPHIA和EUMODIC以及随后的国际小鼠表型分析联盟）中建立的用于监测基因敲除突变小鼠表型的系统筛选。我们在此还描述了基于精子DNA分析和育种研究的定量PCR（qPCR）检测生殖系突变的分析程序。
