Cellular commitment to mortality: analyses of mouse early embryo

细胞对死亡的承诺：小鼠早期胚胎的分析

• 批准号: 7592028
• 负责人: Minoru S Ko
• 金额: $ 62.37万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7592028
• 关键词: Aging; Cells; Cellular biology; Classification; DNA Microarray Chip; DNA Microarray format; Development; Embryo; Fertilization; Gene Activation; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Genomics; Goals; In Situ Hybridization; Molecular; Molecular Profiling; Mus; Names; Numbers; Organ; Pattern; Play; RNA; RNA Degradation; Reverse Transcriptase Polymerase Chain Reaction; Role; Source; Staging; Standards of Weights and Measures; Stem cells; Techniques; Therapeutic; Tissues; base; blastocyst; embryonic stem cell; gene function; implantation; mortality; nuclear reprogramming

We carried out DNA microarray-based global expression profiling of all preimplantation stages in mouse, which revealed and characterized the distinctive patterns of maternal RNA degradation and two major transient waves of de novo transcription: zygotic genome activation (ZGA) and mid-preimplantation gene activation (MGA). Through these analyses, we identified a number of genes that show unique expression patterns. We have demonstrated that one of them, named Zscan4, is expressed exclusively in 2-cell mouse embryos and ES cells and plays a critical role in the progression of preimplantation development. We also found that a gene called Chuk shows constant RNA levels throughout preimplantation development and can be used as an internal standard suitable for quantitative RT-PCR. We also developed a technique to do large-scale Whole Mount In Situ Hybridization (WISH), which allows us to reveal the spatial expression patterns of 91 genes in mouse preimplantation embryos. We are currently studying the functions of these genes in details.

我们进行了基于DNA微阵列的全球表达谱的所有植入前阶段的小鼠，揭示和特点的独特模式的母体RNA降解和两个主要的瞬时波的从头转录：合子基因组激活（ZGA）和中期植入前基因激活（MGA）。通过这些分析，我们确定了一些显示独特表达模式的基因。我们已经证明，其中一个，命名为Zscan 4，只在2-细胞小鼠胚胎和ES细胞中表达，并在植入前发育的进展中起着关键作用。我们还发现，一个名为Chuk的基因在整个着床前发育过程中显示出恒定的RNA水平，可以用作定量RT-PCR的内标。我们还开发了一种技术，做大规模的整体原位杂交（WISH），这使我们能够揭示小鼠植入前胚胎中91个基因的空间表达模式。我们目前正在详细研究这些基因的功能。