Knockout of DNase1l1l abrogates lens denucleation process and causes cataract in zebrafish

Zhang, J., Cui, W.W., Du, C., Huang, Y., Pi, X., Guo, W., Wang, J., Huang, W., Chen, D., Li, J., Li, H., Zhang, J., Ma, Y., Mu, H., Zhang, S., Liu, M., Cui, X., Hu, Y.
Biochimica et biophysica acta. Molecular basis of disease   1866: 165724 (Journal)
Registered Authors
Cui, Xiukun, Liu, Mugen
Cataract, DNase1l1l, Hsf4, Lens denucleation, Zebrafish
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • CRISPR-Cas Systems/genetics
  • Cataract/genetics*
  • Cataract/pathology
  • Cell Nucleus/metabolism
  • Deoxyribonucleases/genetics*
  • Deoxyribonucleases/metabolism
  • Disease Models, Animal
  • Embryo, Nonmammalian
  • Epithelial Cells/cytology
  • Epithelial Cells/metabolism
  • Female
  • Gene Expression Regulation, Developmental*
  • Gene Knockout Techniques
  • Heat Shock Transcription Factors/genetics
  • Heat Shock Transcription Factors/metabolism*
  • Humans
  • Lens, Crystalline/cytology
  • Lens, Crystalline/embryology*
  • Lens, Crystalline/metabolism
  • Lens, Crystalline/pathology
  • Male
  • Zebrafish
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism
32061775 Full text @ BBA Molecular Basis of Disease
Removal of nuclei in lens fiber cells is required for organelle-free zone (OFZ) formation during lens development. Defect in degradation of nuclear DNA leads to cataract formation. DNase2β degrades nuclear DNA of lens fiber cells during lens differentiation in mouse. Hsf4 is the principal heat shock transcription factor in lens and facilitates the lens differentiation. Knockout of Hsf4 in mouse and zebrafish resulted in lens developmental defect that was characterized by retaining of nuclei in lens fiber cells. In previous in vitro studies, we found that Hsf4 promoted DNase2β expression in human and mouse lens epithelial cells. In this study, it was found that, instead of DNase2β, DNase1l1l is uniquely expressed in zebrafish lens and was absent in Hsf4-/- zebrafish lens. Using CRISPR-Cas9 technology, a DNase1l1l knockout zebrafish line was constructed, which developed cataract. Deletion of DNase1l1l totally abrogated lens primary and secondary fiber cell denucleation process, whereas had little effect on the clearance of other organelles. The transcriptional regulation of DNase1l1l was dramatically impaired in Hsf4-/- zebrafish lens. Rescue of DNase1l1l mRNA into Hsf4-/- zebrafish embryos alleviated its defect in lens fiber cell denucleation. Our results in vivo demonstrated that DNase1l1l is the primary DNase responsible for nuclear DNA degradation in lens fiber cells, and Hsf4 can transcriptionally activate DNase1l1l expression in zebrafish.
Genes / Markers
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Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes