Using Functional Genetics in Haploid Cells for Drug Target Identification

Methods Mol Biol. 2019;1953:3-21. doi: 10.1007/978-1-4939-9145-7_1.


Pooled genetic screens are a powerful tool to identify targets for drug development as well as chemogenetic interactions. Various complementary methods for mutagenesis are available to generate highly complex cell populations, including mRNA knockdown, directed genome editing, as well as random genome mutagenesis. With the availability of a growing number of haploid mammalian cell lines, random mutagenesis is becoming increasingly powerful and represents an attractive alternative, e.g., to CRISPR-based screening. This chapter provides a step-by-step protocol for performing haploid gene trap screens.

Keywords: Functional genomics; Gene trap; Genetic screen; Haploid; Stem cell.

MeSH terms

  • Animals
  • Cell Culture Techniques / methods
  • Cell Line
  • Drug Discovery / methods*
  • Drug Evaluation, Preclinical / methods*
  • Flow Cytometry / methods
  • Genetic Testing
  • Haploidy
  • Humans
  • Mice
  • Mutagenesis*
  • Polymerase Chain Reaction / methods
  • Stem Cells / drug effects
  • Stem Cells / metabolism*