Generation and characterization of tamoxifen-inducible Pax9-CreER knock-in mice using CrispR/Cas9

Genesis. 2016 Sep;54(9):490-6. doi: 10.1002/dvg.22956. Epub 2016 Jul 26.

Abstract

Pax9 encodes a paired-box homeodomain (Pax) transcription factor and is critical for the development of multiple organs. Using CrispR/Cas9-mediated homologous directed repair (HDR), we generated a new Pax9-CreER knock-in mouse line in which the CreER(T2) fusion protein is produced after synthesis of endogenous Pax9 protein. We found that tdTomato reporter expression in Pax9-CreER;tdTomato reporter mice is detectable in a similar pattern to the endogenous Pax9 expression, faithfully recapitulating the Pax9 expression domains throughout the embryo and in the adult mouse. At early embryonic stages, the tdTomato reporter is expressed first in the pharyngeal pouch region and later in the craniofacial mesenchyme, somites, limbs, and lingual papillae in the adult tongue. These results demonstrate that this new Pax9-CreER knock-in mouse line can be used for lineage tracing and genetic targeting of Pax9-expressing cells and their progeny in a temporally and spatially controlled manner during development and organogenesis.

Keywords: Pax9; Pax9-CreER; genetic labeling; mouse development.

Publication types

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

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • Gene Knock-In Techniques / methods*
  • Integrases / genetics
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mice
  • Mice, Inbred C57BL
  • PAX9 Transcription Factor / genetics
  • Red Fluorescent Protein
  • Tamoxifen / pharmacology
  • Transcriptional Activation / drug effects

Substances

  • Luminescent Proteins
  • PAX9 Transcription Factor
  • Tamoxifen
  • Cre recombinase
  • Integrases