Gene trapping for drug discovery

Background

Gene trap mutagenesis can be used as an objective, economic and high-throughput tool to dissect genetic influences on cell function and drug response/toxicity. Gene trapping is already established as a means to introduce permanent loss-of-function mutations in ES cells for subsequent transgenic mouse strain generation, but has only very recently been applied to in vitro screening.

Specially designed gene trap vectors randomly insert into the genomic sequence and interrupt transcription of the trapped gene. This results in the creation of mutants with scorable phenotypes for screening or selection. Depending on the design of the gene trap experiment these could be:

  • proliferation rate (eg in the presence of a drug)
  • survival (eg in the presence of a drug)
  • migration
  • morphology
  • secretion
  • marker/reporter expression (GFP/luciferase combined with cell sorting/plate reading)

Technology

Researchers can use gene trap mutagenesis to investigate:

  • the cellular mechanisms/toxicity of a drug (eg lithium as a mood stabiliser)
  • the genes modifying a diseased gene’s effect – rescue/complementation
  • resistance to infection – we have identified mutants that confer resistance to Shigella infection and are exploring the mutants conferring resistance to Toxoplasma infection
  • resistance to inflammation – we are exploring the mutants that alter interleukin/cytokine secretion after pro-inflammatory stimuli
  • metabolic mutants

Key benefits

These include:

  • it can be used a primary identification screen for mutations producing a defined cellular phenotypic assay outcome. Thus target identification is a clear purpose, but with the reassurance of no off-target effects and true causation
  • a rapid and economic alternative to the costly and hard-to-reproduce RNAi library screens for functional genetics or to the HAP1 haploid library that will have many lethal mutations missing

Markets & application

  • academic: exploration of functional biology of cells tailored to research interest
  • commercial: pharmacogenomics, off-target drug effects, toxicity mechanisms

Licensing & development

We are seeking partners for development.