Thompson E M, Adenot P, Tsuji F I, Renard J P
Unité de Biologie du Développement, Institut National de la Recherche Agronomique, Jouy-en Josas, France.
Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1317-21. doi: 10.1073/pnas.92.5.1317.
Study of gene expression kinetics during preimplantation mammalian development is difficult because of the limited amount of material and the usually destructive, static nature of molecular analyses. We describe continuous, noninvasive monitoring of gene expression in preimplantation embryos by using a secreted luminescent reporter, Vargula luciferase. Transgene expression profiles were followed by assaying aliquots of culture medium or by direct visualization of Vargula luciferase secretion from living embryos in real time through photon imaging. With this approach, it is possible to observe epigenetic modulations of gene expression and to link this over time to the developmental capacity of individual embryos. In addition, by developing a strategy where expression from integrated transgenes is enhanced relative to that from nonintegrated DNA, we provide evidence that rapid detection of transgene integration prior to the blastocyst stage should be possible. Thus, imaging of Vargula luciferase secretion may also be useful in the early screening of embryos, for example, in the production of transgenic livestock.
由于材料数量有限以及分子分析通常具有破坏性和静态性，对哺乳动物植入前发育过程中的基因表达动力学进行研究颇具难度。我们描述了通过使用一种分泌型发光报告基因——海萤荧光素酶，对植入前胚胎中的基因表达进行连续、非侵入性监测。通过检测培养基的等分试样或通过光子成像实时直接观察活胚胎分泌的海萤荧光素酶，来追踪转基因表达谱。采用这种方法，有可能观察到基因表达的表观遗传调控，并随着时间的推移将其与单个胚胎的发育能力联系起来。此外，通过开发一种相对于非整合DNA增强整合转基因表达的策略，我们提供了证据表明在囊胚期之前快速检测转基因整合应该是可行的。因此，海萤荧光素酶分泌成像在胚胎的早期筛选中也可能有用，例如在转基因家畜的生产中。
