DUX4 is a multifunctional factor priming human embryonic genome activation

Transient DUX4 expression in human embryonic stem cells induces blastomere-like expression program that is marked by SLC34A2

“…We added two earlier time points before 24 h because the expression of DUX4 target genes might peak earlier, given their temporal expression in the embryo ( De Iaco et al., 2017 ; Hendrickson et al., 2017 ). Here, we confirmed that DUX4 protein was highly expressed already at 6 h after induction but rapidly reduced and disappeared at 48 h ( Figure 2 B), which mimics its dynamics in the embryo ( Vuoristo et al., 2022 ). After filtering out low-quality cells, 65,460 cells were retained for downstream analyses ( Table S2 ).…”

“…Notably, cells at 24 h after 15 and 30 min of treatment showed highly similar expression profiles (r = 0.96; Spearman correlation) ( Figure S1 C). The expression changes of DUX4 target genes ( Table S1 ; Resnick et al., 2019 ) were similar between continuously (4 h) treated cells ( Vuoristo et al., 2022 ) and pulsed cells at 24 h post-treatment, suggesting that the 15- and 30-min pulses were sufficient to activate DUX4 target genes ( Figures 1 D and S1 D). Furthermore, cleavage-stage-specific repetitive elements, such as MLT2A1, MLT2A2, and HERVL, activated by DUX4 ( Geng et al., 2012 ; Hendrickson et al., 2017 ; Young et al., 2013 ), were significantly upregulated at 24 h after pulse ( Figure S1 E).…”

“…DUX4-TetOn hESCs ( Vuoristo et al., 2022 ) were maintained on hESC-qualified Geltrex (Thermo Fisher Scientific) coated tissue culture dishes in Essential 8 culture medium (Thermo Fisher Scientific) in 5% CO 2 at 37°C. The cells were passaged every 3–5 days after a 3-min incubation with 0.5 mM EDTA (Thermo Fisher Scientific).…”

“…Two samples collected immediately after induction were excluded due to a low number of mapped reads. The STRT RNA-seq data of continuous DUX4 induction, treated by doxycycline for 4 h, were obtained from Vuoristo et al. (2022) and reprocessed as described elsewhere ( https://github.com/my0916/STRT2 ; Ezer et al., 2021 ).…”

“…DUX4 , a double homeobox transcription factor, is transiently expressed in the human cleavage stage embryo ( Töhönen et al., 2017 ) and regulates human EGA by inducing transcription of cleavage-stage-specific genes and repetitive elements ( De Iaco et al., 2017 ; Hendrickson et al., 2017 ; Vuoristo et al., 2022 ; Whiddon et al., 2017 ). However, DUX4 overexpression in somatic cells leads to cell death both in vitro and in vivo ( Bosnakovski et al., 2008 ; Kowaljow et al., 2007 ; Rickard et al., 2015 ; Wallace et al., 2011 ).…”