A reversible gene trap collection empowers haploid genetics in human cells.

Journal article

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.

Authors: Bürckstümmer, T; Banning, C; Hainzl, P; Schobesberger, R; Kerzendorfer, C

• Journal: Nature methods
• Volume: 10
• Issue: 10
• Pages: 965-971
• Publication date: 2013-10-01
• DOI: 10.1038/nmeth.2609
• EISSN: 1548-7105
• ISSN: 1548-7091
• Language: English
• Keywords: Humans; Reverse Genetics; Molecular Sequence Data; Cell Line, Tumor; Genome, Human; Haploidy; Mutagenesis, Insertional; Gene Library