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首页> 外文期刊>Nucleus >Positional changes of a pluripotency marker gene during structural reorganization of fibroblast nuclei in cloned early bovine embryos
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Positional changes of a pluripotency marker gene during structural reorganization of fibroblast nuclei in cloned early bovine embryos

机译：克隆的早期牛胚胎中成纤维细胞核结构重组过程中多能性标记基因的位置变化

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Cloned bovine preimplantation embryos were generated by somatic cell nuclear transfer (SCNT) of bovine fetal fibroblasts with a silent copy of the pluripotency reporter gene GOF , integrated at a single site of a chromosome 13. GOF combines the regulatory Oct4/Pou5f1 sequence with the coding sequence for EGFP. EGFP expression served as a marker for pluripotency gene activation and was consistently detected in preimplantation embryos with 9 and more cells. Three-dimensional radial nuclear positions of GOF , its carrier chromosome territory and non-carrier homolog were measured in nuclei of fibroblasts, and of day 2 and day 4 embryos, carrying 2 to 9 and 15 to 22 cells, respectively. We tested, whether transcriptional activation was correlated with repositioning of GOF toward the nuclear interior either with a corresponding movement of its carrier chromosome territory 13 or via the formation of a giant chromatin loop. A significant shift of GOF away from the nuclear periphery was observed in day 2 embryos together with both carrier and non-carrier chromosome territories. At day 4, GOF , its carrier chromosome territory 13 and the non-carrier homolog had moved back toward the nuclear periphery. Similar movements of both chromosome territories ruled out a specific GOF effect. Pluripotency gene activation was preceded by a transient, radial shift of GOF toward the nuclear interior. The persistent co-localization of GOF with its carrier chromosome territory rules out the formation of a giant chromatin loop during GOF activation.
机译：牛胎成纤维细胞的体细胞核移植（SCNT）具有多能性报告基因 GOF的无声拷贝，并整合在13号染色体的单个位点上，从而产生了克隆的牛植入前胚胎。 i> Oct4 / Pou5f1序列以及EGFP的编码序列。 EGFP表达可作为多能性基因激活的标记，并且在植入前的胚胎中有9个及以上的细胞中始终检测到EGFP的表达。在成纤维细胞的核中以及分别携带2至9个和15至22个细胞的第2天和第4天胚胎中，测量了GOF的三维放射状核位置，其载体染色体区域和非载体同源物。我们测试了转录激活是否与GOF朝向核内部的重新定位有关，要么与它的载体染色体区域13的相应移动有关，要么与巨染色质环的形成有关。在第2天的胚胎以及携带者和非携带者的染色体区域中都观察到了GOF离开核外围的显着变化。在第4天，GOF的携带者染色体区域13和非携带者同源物已移回核外围。两个染色体区域的相似运动排除了特定的GOF效应。多能性基因激活之前，GOF向核内部发生了短暂的径向移动。 GOF及其载体染色体区域的持续共定位排除了GOF激活期间巨染色质环的形成。

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