Transcription inhibition in early mouse embryos impacts transcriptome at 4-cell stage

Mouse zygotes were treated with DRB to inhibit transcription activity, and early embryos were collected at 4-cell stage to examine transcriptome alteration by RNA-seq. We treated mouse zygotes 6 h post-fertilization using KSOM medium supplemented with 20 μM of DRB (5,6 - Dichloro - 1 - β - D - ribofuranosylbenzimidazole), a transcription inhibitor. Following a 20-hour incubation period with DRB, the embryos were meticulously washed to ensure complete removal of the inhibitor and then transferred to fresh KSOM medium. The culturing process was extended for an additional 26 hours, and subsequently, 4-cell embryos from both groups were collected and subjected to transcriptome sequencing using RNA-seq to examine the alterations in the transcriptome.

本研究旨在通过RNA-seq检测4-细胞阶段早期胚胎的转录组变化，探究转录抑制剂DRB（5,6-二氯-1-β-D-呋喃核糖基苯并咪唑，5,6-Dichloro-1-β-D-ribofuranosylbenzimidazole）对小鼠受精卵转录活性的影响。具体实验方案如下：于小鼠受精后6小时，采用添加了20 μM DRB的KSOM培养基处理受精卵，DRB为一种经典的转录抑制剂。经DRB孵育20小时后，对胚胎进行细致清洗以完全去除抑制剂，随后将胚胎转移至新鲜KSOM培养基中继续培养。继续培养26小时后，收集两组的4-细胞胚胎，通过RNA-seq进行转录组测序以分析转录组的改变情况。