Characterization and function of transcriptional elongation during zygotic genome activation in mouse embryos [ChIL-seq]. Characterization and function of transcriptional elongation during zygotic genome activation in mouse embryos [ChIL-seq]

The work associated to these datasets focuses on the investigation of transcriptional regulation during the earliest stages of mammalian embryogenesis. In particular, ChILseq and RNAseq datasets were generated to study the process of zygotic genome activation in mouse embryos. After fertilisation, the mouse embryo activates its own genome for the first time in a process known as zygotic genome activation, which occurs primarily on two main waves. The first wave occurs in zygotes, and the second one at the late 2-cell stage. To understand how the RNA Polymerase II functions during these waves, we have implemented a ChILseq approach to generate genome-wide maps of Ser5 and Ser2 phosphorylated forms of RNA Polymerase II in zygotes, 2- and 8-cell stage embryos. In addition, we have investigated how elongation factors, such as NELF and DSIF regulate major ZGA by performing single embryo RNAseq using SMART-seq2 in 2-cell stage embryos upon depletion of Spt5, NELFE and upon chemical inhibition of Cdk9. Overall design: ChIL-seq experiments using Pol II Ser5P, Ser2P and IgG antibody in 3 biological replicates

本数据集相关的研究工作聚焦于哺乳动物胚胎发生早期阶段的转录调控机制探究。具体而言，我们生成了ChIL测序（ChILseq）与RNA测序（RNAseq）数据集，以研究小鼠胚胎中的合子基因组激活（zygotic genome activation, ZGA）过程。受精后，小鼠胚胎会首次激活自身基因组，该过程即合子基因组激活，主要分为两个波次：第一波发生于受精卵阶段，第二波则出现于晚期2细胞期。为阐明RNA聚合酶II（RNA Polymerase II）在上述波次中的功能机制，我们采用ChILseq技术，绘制了受精卵、2细胞期及8细胞期胚胎中RNA聚合酶II的Ser5与Ser2磷酸化形式的全基因组结合图谱。此外，我们对经Spt5、NELFE敲降以及Cdk9化学抑制处理的2细胞期胚胎开展基于SMART-seq2的单胚胎RNA测序，以此探究NELF与DSIF等转录延伸因子对主要合子基因组激活的调控作用。整体实验设计：针对Pol II Ser5P、Ser2P及IgG抗体，设置3次生物学重复开展ChIL-seq实验。