NicE-seq in Suv39h2 knockdown 8-cell stage

Determination of accessible chromatin regions in the 8-cell stage mouse embryo after Suv39h2 knockdown The study aimed to address a potential function of H3K9me3 in early mouse development by assessing its impact on chromatin compaction. The histone methyltransferase responsible for de novo H3K9me3 at fertilization Suv39h2, was knocked down by microinjection of dsRNA targeting Suv39h2 in the early zygote. Embryos were then cultured until the 8-cell stage of development. They were then fixed and chromatin compaction was assessed by NicE-seq in pools of 10 8-cell stage embryos. Overall design: Mouse zygotes were microinjected with double stranded RNA (approx 400bp) targeting N-terminal Suv39h2 or Lacz as control. They were cultured until the 8-cell stage and pools of 10 were fixed in 1% PFA. NicE-seq (nicking enzyme assisted sequencing) was applied accoridng to Ponnaluri et al., 2017.