Haplogen GmbH, Vienna, Austria.
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.
敲除细胞集是研究酵母等模式生物的宝贵工具。然而，目前还没有人类细胞的大规模敲除细胞集。我们利用近单倍体人类细胞中的基因捕获诱变，建立了一个生成和分离单个“基因捕获细胞”的平台，并利用该平台制备了携带单个基因捕获插入的人类细胞系集。在大多数情况下，插入可以逆转。这个不断增长的文库涵盖了 3396 个基因，占表达基因组的三分之一，它带有 DNA 条形码，可进行各种细胞表型的系统筛选。我们研究了细胞对 TNF-α、TGF-β、IFN-γ 和 TNF 相关凋亡诱导配体 (TRAIL) 的反应，以此来说明这个独特的同源人类细胞系集的价值。
