A reversible gene trap collection empowers haploid genetics in human cells.
Bürckstümmer T., Banning C., Hainzl P., Schobesberger R., Kerzendorfer C., Pauler FM., Chen D., Them N., Schischlik F., Rebsamen M., Smida M., Fece de la Cruz F., Lapao A., Liszt M., Eizinger B., Guenzl PM., Blomen VA., Konopka T., Gapp B., Parapatics K., Maier B., Stöckl J., Fischl W., Salic S., Taba Casari MR., Knapp S., Bennett KL., Bock C., Colinge J., Kralovics R., Ammerer G., Casari G., Brummelkamp TR., Superti-Furga G., Nijman SMB.
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.