Single cell RNA-seq Analysis of Wild Type(WT), Dux knockout(KO) and Dux overexpression(OE) embryos

We tested the transcriptome of embryos from WT and Dux KO mice including zygotes (hCG 28h), early 2-cell (hCG 31-32h), middle 2-cell(hCG 41-42h), late 2-cell(hCG 47-48h) and, Dux mRNA injected embryos including zygote(5h after injection, hCG 28h), early 4-cell(6h after injection, hCG 54h) and late 4-cell (17h after injection, hCG 65h) using the Covaris DNA shearing protocol for Smart-seq sequence library generation. We found that the activation of Dux is important but not essential for ZGA, but the silencing and elimination of Dux is strictly necessary for early embryonic development. Embryos mRNA profiles of WT,Dux KO and Dux OE were generated by deep sequencing, using Illumina NovaSeq 6000.