1.Institute of Stem Cell and Regenerative Biology, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan, China
2.Frontiers Science Center for Animal Breeding and Sustainable Production, Huazhong Agricultural University, Ministry of Education, Wuhan, China
Abstract Text: Emerging evidence highlights the regulatory role of PRD-like homeobox transcription factors (TFs) in embryonic genome activation (EGA). However, the majority of PRD-like genes are lost in rodents, thus urging an investigation for PRD-like TFs in non-rodent mammals. Here we showed that PRD-like TFs, such as ARGFX, DPRX, DUXB, LEUTX, TPRX, were transiently expressed during EGA in non-rodent fertilized embryos, including human, monkey, and pig, yet all these TFs exhibited inadequate expression in their cloned embryos. In this study, applying pig as the research model, we identified LEUTX as a key PRD-like activator of porcine EGA through reporter selection assay, combined RNA-seq data analysis with LEUTX genomic profiling via CUT&Tag. Furthermore, we found that LEUTX overexpression restored EGA failure (reactivated 76% EGA genes), improved blastocyst formation (elevated from 13.9% to 26.2%) and cloning efficiency (elevated from 0.75% to 4.11%) in porcine cloned embryos. Mechanistically, by conducting co-IP and proximity ligation assay, we revealed that LEUTX opened EGA-related genomic regions and established histone acetylation via recruiting acetyltransferases p300 and KAT2A. Utilizing PRD-like TFs' DNA-binding motif, we also developed a reporter system for real-time EGA monitoring to select early embryos with good quality. These findings highlight LEUTX as a conserved PRD-like TF governing non-rodent mammalian EGA, which also benefits somatic cell reprogramming in these mammals.
