| Products/Services Used | Details | Operation |
|---|---|---|
| Peptide Synthesis | Promega Cat# E2311 RiboLock Thermo Fisher Scientific Cat# EO0382 MMLV-RTase Promega Cat# M1701 DUX4 9aaTAD peptide for binding analysis: CGLLLDELLASPEFLQQ GenScript N/A DUX4 KBM peptide for binding analysis: EEEYRALLEE GenScript N/A Histrap HP column (1 ml) GE Healthcare Cat# 17524701 Resource Q column (1 ml) GE -RTase Promega Cat# M1701 DUX4 9aaTAD peptide for binding analysis: CGLLLDELLASPEFLQQ GenScript N/A DUX4 KBM peptide for binding analysis: EEEYRALLEE GenScript N/A Histrap HP column (1 ml) GE Healthcare Cat# 17524701 Resource Q column (1 ml) GE Healthcare Cat# 17117701 Superdex 75 10/300 GL column GE Healthcare ) and KBM ( 416 EYRALL 421 ) peptides to KIX; the homeodomain of human LEUTX with His-Tag was used as a negative control. Peptides were ordered from GenScript ( key resources table ) and dissolved in deionized water. The final concentration of KIX in the assay was 20 nM, and the concentration of each peptide -tight-hMAFA-ires-EmGFP-pA-PGK-Puro vector (a kind gift from Dr. Diego Balboa, Stem Cells and Metabolism Research Program, University of Helsinki) at GenScript (Genscript, NJ, USA). For the dCas9-DUX4-C construct, the C-terminal part of the DUX4 was PCR amplified from the pB-tight-hDUX4-ires-EmGFP-pA-PGK-Puro -hMAFA-ires-EmGFP-pA-PGK-Puro vector (a kind gift from Dr. Diego Balboa, Stem Cells and Metabolism Research Program, University of Helsinki) at GenScript (Genscript, NJ, USA). For the dCas9-DUX4-C construct, the C-terminal part of the DUX4 was PCR amplified from the pB-tight-hDUX4-ires-EmGFP-pA-PGK-Puro vector (using | Get A Quote |
SummaryDouble homeobox 4 ( DUX4 ) is expressed at the early pre-implantation stage in human embryos. Here we show that induced human DUX4 expression substantially alters the chromatin accessibility of non-coding DNA and activates thousands of newly identified transcribed enhancer-like regions, preferentially located within ERVL-MaLR repeat elements. CRISPR activation of transcribed enhancers by C-terminal DUX4 motifs results in the increased expression of target embryonic genome activation (EGA) genes ZSCAN4 and KHDC1P1 . We show that DUX4 is markedly enriched in human zygotes, followed by intense nuclear DUX4 localization preceding and coinciding with minor EGA. DUX4 knockdown in human zygotes led to changes i... More
