ENU-based phenotype-driven screening

Methods Mol Biol. 2012:844:1-13. doi: 10.1007/978-1-61779-527-5_1.

Abstract

Deciphering the contribution of individual genes and in turn pathways to cellular processes can be complicated and is often based on prior knowledge or assumptions of gene function. Phenotype-driven mutagenesis screens based around n-ethyl-n-nitrosurea (ENU) have been successful in a wide range of physiological systems in identifying novel genes that contribute to a given phenotype. Here, we describe methodologies we have employed in analysing cellular phenotypes in pipelines of mutagenised mice. Examples of primary screens to identify outliers, and secondary screens to provide a more detailed characterisation are outlined.

MeSH terms

  • Animals
  • Blood Cell Count / instrumentation
  • Blood Cell Count / methods
  • Bone Marrow Cells / metabolism
  • Breeding
  • Cytokines / analysis
  • Cytokines / metabolism
  • Ethylnitrosourea / administration & dosage
  • Ethylnitrosourea / toxicity*
  • Female
  • Flow Cytometry / methods
  • Leukocytes / drug effects*
  • Leukocytes / metabolism
  • Macrophages / drug effects
  • Macrophages / metabolism
  • Male
  • Mice
  • Mice, Inbred C3H
  • Mice, Inbred C57BL
  • Mutagenesis*
  • Phenotype*

Substances

  • Cytokines
  • Ethylnitrosourea