Wdhd1 is essential for early mouse embryogenesis

Hsiang-Hsuan Fan; Kuo-Hong Lee; You-Tzung Chen; Li-Jyuan Lin; Tsung-Lin Yang; Shu-Wha Lin; I-Shing Yu

doi:10.1016/j.bbamcr.2021.119011

Wdhd1 is essential for early mouse embryogenesis

Biochim Biophys Acta Mol Cell Res. 2021 May;1868(6):119011. doi: 10.1016/j.bbamcr.2021.119011. Epub 2021 Mar 11.

Authors

Hsiang-Hsuan Fan¹, Kuo-Hong Lee¹, You-Tzung Chen², Li-Jyuan Lin³, Tsung-Lin Yang⁴, Shu-Wha Lin⁵, I-Shing Yu⁶

Affiliations

¹ Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan University, Taipei 100, Taiwan.
² Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan University, Taipei 100, Taiwan; Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei 100, Taiwan; NTU Centers of Genomic and Precision Medicine, National Taiwan University, Taipei 100, Taiwan; Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei 100, Taiwan; Center for Biotechnology, National Taiwan University, Taipei 100, Taiwan.
³ NTU Centers of Genomic and Precision Medicine, National Taiwan University, Taipei 100, Taiwan.
⁴ Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei 100, Taiwan; Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei 100, Taiwan; Department of Otolaryngology, National Taiwan University Hospital, Taipei 100, Taiwan.
⁵ NTU Centers of Genomic and Precision Medicine, National Taiwan University, Taipei 100, Taiwan; Department of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan University, Taipei 100, Taiwan.
⁶ NTU Centers of Genomic and Precision Medicine, National Taiwan University, Taipei 100, Taiwan; Laboratory Animal Center, College of Medicine, National Taiwan University, Taipei 100, Taiwan. Electronic address: isyu@ntu.edu.tw.

PMID: 33716172
DOI: 10.1016/j.bbamcr.2021.119011

Abstract

WD repeat and HMG-box DNA binding protein 1 (Wdhd1) is the mouse ortholog of budding yeast Chromosome Transmission Fidelity 4 (CTF4), the protein product of which integrates the MCM2-7 helicase and DNA polymerase α/primase complex to initiate DNA replication. Previous work in fruit flies, Xenopus egg extracts, and human cell lines suggest that Wdhd1 is required for efficient DNA synthesis. However, rigorous in vivo functional studies on Wdhd1 in mammals are unavailable. In the present study, we have successfully generated a Wdhd1 null allele in mice through CRISPR/Cas9-mediated genome editing to investigate the role of Wdhd1 in embryogenesis in vivo. We characterized Wdhd1 expression using quantitative reverse-transcription polymerase chain reaction, and assessed embryonic cell proliferation by histology in both pre- and peri-implantation embryos. While Wdhd1 heterozygous mutant mice were grossly normal and fertile, we observed a reduction in cell proliferation by the gastrulation stage in Wdhd1 homozygous null mutant embryos which severely hampered their growth and viability. These results indicate that Wdhd1 plays a major role in cell proliferation during embryogenesis in mice.

Keywords: CRISPR/Cas9; DNA replication; Embryogenesis; Mouse; Proliferation; Wdhd1.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Cas Systems
Cell Culture Techniques
Cell Line
Cell Proliferation
DNA-Binding Proteins / genetics*
DNA-Binding Proteins / metabolism*
Embryonic Development*
Fertility
Gastrulation
Gene Editing
Loss of Function Mutation
Mice

Substances

DNA-Binding Proteins