Tools and Techniques for Wt1-Based Lineage Tracing

Methods Mol Biol. 2016:1467:41-59. doi: 10.1007/978-1-4939-4023-3_4.

Abstract

The spatiotemporal expression pattern of Wt1 has been extensively studied in a number of animal models to establish its function and the developmental fate of the cells expressing this gene. In this chapter, we review the available animal models for Wt1-expressing cell lineage analysis, including direct Wt1 expression reporters and systems for permanent Wt1 lineage tracing. We describe the presently used constitutive or inducible genetic lineage tracing approaches based on the Cre/loxP system utilizing Cre recombinase expression under control of a Wt1 promoter.To make these systems accessible, we provide laboratory protocols that include dissection and processing of the tissues for immunofluorescence and histopathological analysis of the lineage-labeled Wt1-derived cells within the embryo/tissue context.

Keywords: Cell lineage tracing; Cre-ERT2; Cre/loxP; Inducible Cre recombinase; Rosa26 reporter mice; Tamoxifen; Wt1.

Publication types

  • Review

MeSH terms

  • Animals
  • Cell Lineage
  • Cell Tracking / methods*
  • Genes, Reporter
  • Integrases / genetics
  • Integrases / metabolism
  • Mice
  • Mice, Transgenic
  • Models, Animal
  • Promoter Regions, Genetic
  • Repressor Proteins / genetics*
  • Repressor Proteins / metabolism*
  • Tamoxifen / pharmacology
  • WT1 Proteins

Substances

  • Repressor Proteins
  • WT1 Proteins
  • WT1 protein, mouse
  • Tamoxifen
  • Cre recombinase
  • Integrases