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SRX17893697: GSM6641844: RNA_WT_L2C_rep1; Mus musculus; RNA-Seq
1 ILLUMINA (Illumina NovaSeq 6000) run: 3.5M spots, 1.1G bases, 333.7Mb downloads

External Id: GSM6641844_r1
Submitted by: Tsinghua University
Study: OBOX regulates murine zygotic genome activation and early development
show Abstracthide Abstract
Zygotic genome activation (ZGA) activates the quiescent genome to enable the maternal-to-zygotic transition. However, the identity of transcription factors (TFs) that underlie mammalian ZGA in vivo remains unresolved. Here, we showed that OBOX, a PRD-like homeobox domain TF family that includes 66 members (OBOX1-8), are key regulators of mouse ZGA. Knockout mice deficient for maternally transcribed Obox1/2/5/7 and zygotically expressed Obox3/4 led to 2-4 cell arrest accompanied by impaired ZGA. Maternal and zygotic expressed OBOX redundantly support embryonic development as Obox KO defects can be rescued by restoring either of them. Chromatin binding analysis revealed Obox knockout preferentially affected OBOX-binding targets. Mechanistically, OBOX facilitated RNA Pol II ?pre-configuration?, as Pol II relocates from the initial 1-cell binding targets to ZGA gene promoters and distal enhancers. The impaired Pol II pre-configuration in Obox mutants was accompanied by downregulation of ZGA genes, chromatin accessibility transition defects, as well as aberrant activation of one-cell Pol II targets. Finally, OBOX ectopically activated ZGA genes and MERVL in mESCs. Hence, these data demonstrate that OBOX regulates murine ZGA and early embryogenesis. Overall design: By employing RNA-seq and Pol II Stacc-seq/ATAC-Seq in Obox mzKO embryos as well as RNA-seq in Obox OE mESCs, we revealed maternal OBOX is the key transcription factor regulating murine zygotic genome activation.
Sample: RNA_WT_L2C_rep1
SAMN31286211 • SRS15410894 • All experiments • All runs
Organism: Mus musculus
Library:
Name: GSM6641844
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: For embryos, the zona pellucida was gently removed by treatment with Tyrode's solution (Sigma, T1844). Then the embryos were lysed in lysis buffer containing RNase inhibitor. 2i ESCs were lysed in lysis buffer containing RNase inhibitor. The following steps followed the Smart-seq2 protocol as previously described (Picelli et al., 2014). For whole genome sequence, the tail tip DNA were extracted with isopropanol precipitation method. The Smart-seq2 libraries of embryos were prepared as previously described (Picelli et al., 2014). Embryos were lysed in lysis buffer containing RNase inhibitor according to the user manual. The library was quantified using Qubit and Agilent 2100 before being subjected to sequencing. For stacc-seq, the library was prepared following the protocol as descripted before (Liu et al., 2020). For ATAC-seq, the library was prepared following the protocol as descripted before (Buenrostro et al., 2013; Wu ET AL., 2016). The Smart-seq2, WGS (Picelli et al., 2014) and stacc-seq (Liu et al., 2020) library strategies were followed the protocols described.
Runs: 1 run, 3.5M spots, 1.1G bases, 333.7Mb
Run# of Spots# of BasesSizePublished
SRR219078153,545,8391.1G333.7Mb2023-04-24

ID:
24827371

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