Generation of a multipurpose Prdm16 mouse allele by targeted gene trapping

Dis Model Mech. 2017 Jul 1;10(7):909-922. doi: 10.1242/dmm.029561. Epub 2017 Apr 19.

Abstract

Gene trap mutagenesis is a powerful tool to create loss-of-function mutations in mice and other model organisms. Modifications of traditional gene trap cassettes, including addition of conditional features in the form of Flip-excision (FlEx) arrays to enable directional gene trap cassette inversions by Cre and Flpe site-specific recombinases, greatly enhanced their experimental potential. By taking advantage of these conditional gene trap cassettes, we developed a generic strategy for generating conditional mutations and validated this strategy in mice carrying a multipurpose allele of the Prdm16 transcription factor gene. We demonstrate that the gene trap insertion creates a null mutation replicating the Pierre Robin sequence-type cleft palate phenotype of other Prdm16 mutant mice. Consecutive breeding to Flpe and Emx1IREScre deleter mice spatially restricted Prdm16 loss to regions of the forebrain expressing the homeobox gene Emx1, demonstrating the utility of the technology for the analysis of tissue-specific gene functions.

Keywords: Cleft palate; Conditional gene trap; Mandible; Micrognathia; Pierre Robin sequence.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles*
  • Animals
  • Brain / metabolism
  • Breeding
  • DNA-Binding Proteins / genetics*
  • Embryo, Mammalian / cytology
  • Embryonic Stem Cells / metabolism
  • Gene Deletion
  • Gene Targeting*
  • Genes, Reporter
  • Genetic Vectors / metabolism
  • Head / embryology
  • Mice
  • Mutation / genetics
  • Organ Specificity
  • Phenotype
  • Transcription Factors / genetics*

Substances

  • DNA-Binding Proteins
  • Prdm16 protein, mouse
  • Transcription Factors