A reversible gene trap collection empowers haploid genetics in human cells

Nat Methods. 2013 Oct;10(10):965-71. doi: 10.1038/nmeth.2609. Epub 2013 Aug 25.


Knockout collections are invaluable tools for studying model organisms such as yeast. However, there are no large-scale knockout collections of human cells. Using gene-trap mutagenesis in near-haploid human cells, we established a platform to generate and isolate individual 'gene-trapped cells' and used it to prepare a collection of human cell lines carrying single gene-trap insertions. In most cases, the insertion can be reversed. This growing library covers 3,396 genes, one-third of the expressed genome, is DNA-barcoded and allows systematic screens for a wide variety of cellular phenotypes. We examined cellular responses to TNF-α, TGF-β, IFN-γ and TNF-related apoptosis-inducing ligand (TRAIL), to illustrate the value of this unique collection of isogenic human cell lines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Gene Library*
  • Genome, Human
  • Haploidy*
  • Humans
  • Molecular Sequence Data
  • Mutagenesis, Insertional / methods*
  • Reverse Genetics / methods*

Associated data

  • GENBANK/KF179301
  • GEO/GSE48848