Using CRISPR/Cas9 engineering to generate a mouse with a conditional knockout allele for the promyelocytic leukemia zinc finger transcription factor

Genesis. 2019 Mar;57(3):e23281. doi: 10.1002/dvg.23281. Epub 2019 Jan 23.

Abstract

The promyelocytic leukemia zinc finger (PLZF) transcription factor mediates a wide-range of biological processes. Accordingly, perturbation of PLZF function results in a myriad of physiologic defects, the most conspicuous of which is abnormal skeletal patterning. Although whole body knockout of Plzf in the mouse (Plzf KO ) has significantly expanded our understanding of Plzf function in vivo, a conditional knockout mouse model that enables tissue or cell-type specific ablation of Plzf has not been developed. Therefore, we used CRISPR/Cas 9 gene editing to generate a mouse model in which exon 2 of the murine Plzf gene is specifically flanked (or floxed) by LoxP sites (Plzf f/f ). Crossing our Plzf f/f mouse with a global cre-driver mouse to generate the Plzf d/d bigenic mouse, we demonstrate that exon 2 of the Plzf gene is ablated in the Plzf d/d bigenic. Similar to the previously reported Plzf KO mouse, the Plzf d/d mouse exhibits a severe defect in skeletal patterning of the hindlimb, indicating that the Plzf f/f mouse functions as designed. Therefore, studies in this short technical report demonstrate that the Plzf f/f mouse will be useful to investigators who wish to explore the role of the Plzf transcription factor in a specific tissue or cell-type.

Keywords: CRISPR/Cas9; conditional allele; cre recombinase; hindlimb; homology directed repair; mouse; polydactyly; promyelocytic leukemia zinc finger; skeletal patterning.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Body Patterning
  • Bone and Bones / abnormalities
  • Bone and Bones / embryology
  • CRISPR-Cas Systems*
  • Gene Knockout Techniques / methods
  • Hindlimb / abnormalities
  • Hindlimb / embryology
  • Mice
  • Mice, Inbred C57BL
  • Promyelocytic Leukemia Zinc Finger Protein / deficiency
  • Promyelocytic Leukemia Zinc Finger Protein / genetics*

Substances

  • Promyelocytic Leukemia Zinc Finger Protein
  • Zbtb16 protein, mouse