Bai Guang-Yu, Song Si-Hang, Sun Rui-Zhen, Zhang Zi-Hui, Li Jingyu, Wang Zhen-Dong, Liu Zhong-Hua, Lei Lei
Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, China.
Laboratory of Embryo Biotechnology, College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
Oncotarget. 2017 Jul 18;8(41):69863-69873. doi: 10.18632/oncotarget.19418. eCollection 2017 Sep 19.
Somatic cell nuclear transfer is an important technique for life science research, but its efficiency is still extremely low, and most genes that are important during early development, such as X chromosome-linked genes, are not appropriately expressed during this process. Poly (ADP-ribose) polymerase (PARP) is an enzyme that transfers ADP ribose clusters to target proteins. PARP family members such as PARP1 participate in cellular signalling pathways through poly (ADP-ribosylation) (PARylation), which ultimately promotes changes in chromatin structure, gene expression, and the localization and activity of proteins that mediate signalling responses. PARP1 is associated with X chromosome inactivation (Xi). Here, we showed that abnormal Xi occurs in somatic cell nuclear transfer (NT) blastocysts, whereas in female blastocysts derived from cumulus cell nuclear transfer, both X chromosomes were inactive. expression was higher in female NT blastocysts than that in intracytoplasmic sperm injection (ICSI) embryos but not in male NT blastocysts. After knocking down expression, both the pre-rRNA 47S and X-inactivation-specific transcript () levels increased. Moreover, the expression of genes on the inactivated X chromosome, such as and were also increased in the NT embryos. However, the development of Parp1si NT embryos was impaired, although total RNA sequencing showed that overall gene expression between the Parp1si NT blastocysts and the control was similar. Our findings demonstrate that increases in the expression of several genes on the X chromosome and of rRNA primary products in NT blastocysts with disrupted expression are insufficient to rescue the impaired development of female cloned mouse embryos and could even exacerbate the associated developmental deficiencies.
体细胞核移植是生命科学研究中的一项重要技术，但其效率仍然极低，并且大多数在早期发育过程中重要的基因，如X染色体连锁基因，在此过程中并未得到适当表达。聚（ADP - 核糖）聚合酶（PARP）是一种将ADP核糖簇转移到靶蛋白的酶。PARP家族成员如PARP1通过聚（ADP - 核糖基化）（PARylation）参与细胞信号通路，最终促进染色质结构、基因表达以及介导信号反应的蛋白质的定位和活性发生变化。PARP1与X染色体失活（Xi）有关。在这里，我们表明体细胞核移植（NT）囊胚中出现异常的Xi，而在来自卵丘细胞核移植的雌性囊胚中，两条X染色体均失活。 表达在雌性NT囊胚中高于胞浆内单精子注射（ICSI）胚胎，但在雄性NT囊胚中并非如此。敲低 表达后，前体rRNA 47S和X失活特异性转录本（ ）水平均升高。此外，NT胚胎中失活X染色体上的基因如 和 的表达也增加。然而，Parp1si NT胚胎的发育受到损害，尽管全RNA测序显示Parp1si NT囊胚与对照之间的整体基因表达相似。我们的研究结果表明，在 表达受到干扰的NT囊胚中，X染色体上几个基因以及rRNA初级产物表达的增加不足以挽救雌性克隆小鼠胚胎受损的发育，甚至可能加剧相关的发育缺陷。
